Technical field

[0001]

　The present invention provides antibodies or fragments thereof against caprin-1, relates to a novel pharmaceutical use as treatment of cancer and / or prophylactic agent.

Background technique

[0002]

　Cancer is a disease that among all of the causes of death, those treatments that are currently practiced in combination with radiation therapy and chemotherapy mainly surgical therapy. Despite the discovery of recent new surgical technique development and new anti-cancer agents, except for some cancers, treatment results of cancers are has not yet been significantly improved. In recent years, advances in molecular biology and cancer immunology, antibodies, which specifically reacts with cancer, cancer antigens recognized by cytotoxic T cells, genes, etc. encoding the cancer antigens have been identified, there is a growing expectation to a specific cancer therapy in which the cancer antigens to target.

[0003]

　Ｃｙｔｏｐｌａｓｍｉｃ－ａｃｔｉｖａｔｉｏｎ　ａｎｄ　ｐｒｏｌｉｆｅｒａｔｉｏｎ－ａｓｓｏｃｉａｔｅｅｄ　ｐｒｏｔｅｉｎ　１（ＣＡＰＲＩＮ－１）は、休止期の正常細胞が活性化や細胞分裂を起こす際に発現し、また細胞内でＲＮＡと細胞内ストレス顆粒を形成してｍＲＮＡの輸送、翻訳の制御に関与することなどが知られている細胞内タンパク質として知られていたが、癌細胞の表面に特異的に発現することが見出され、癌治療のための抗体医薬のターゲットとして研究が進められている（特許文献１～１９）。

CITATION

Patent literature

[0004]

Patent Document 1: WO2010 / 016526
Patent Document 2: WO2011 / 096517
Patent Document 3: WO2011 / 096528
Patent Document 4: WO2011 / 096519
Patent Document 5: WO2011 / 096533
Patent Document 6: WO2011 / 096534
Patent Document 7: WO2011 / 096535
patent Document 8: WO2013 / 018886
Patent Document 9: WO2013 / 018894
Patent Document 10: WO2013 / 018892
Patent Document 11: WO2013 / 018891
Patent Document 12: WO2013 / 018889
Patent Document 13: WO2013 / 018883
Patent Document 14: WO2013 / 125636
Patent Document 15: WO2013 / 125654
Patent Document 16: WO2013 / 125630
Patent Document 17: WO2013 / 125640
Patent Document 18: WO2013 / 147169
Patent Document 19: WO2013 / 147176

Summary of the invention

Problems that the Invention is to Solve

[0005]

　The purpose of the present invention, the caprin-1 expressed specifically on the surface of cancer cells were targeted, than conventional antibodies issues create superior antibody of anti-tumor activity, as a therapeutic and / or prophylactic agent for cancer it is to provide a use.

Means for Solving the Problems

[0006]

　The present invention has the following features.

[0007]

　In the present invention, and a light chain variable region comprising the heavy chain variable region SEQ ID NO: 4, 5 and 6 comprising SEQ ID NO: 1, 2 and 3, and antibodies with caprin-1 protein and immunological reactivity or characterized in that it comprises a fragment thereof as an active ingredient, a pharmaceutical composition for the treatment and / or prevention of cancer.

[0008]

　In that embodiment, the cancer is breast cancer, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer , sarcoma, mast cell tumors, melanoma, adrenocortical cancer, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer, or head and neck cancer.

[0009]

　In another embodiment, the antibody is a human antibody, humanized antibody, chimeric antibody, single chain antibody or a multispecific antibody (e.g. bispecific antibodies).

[0010]

　This specification includes the contents as disclosed in the specification and / or drawings of which is the basis of the present application claims priority of Japanese Patent Application No. 2013-166164.

Effect of the invention

[0011]

　Antibodies to caprin-1 according to the present invention, the failure of cancer cells. Thus, antibodies against caprin-1 according to the present invention are useful for the treatment or prevention of cancer.

DESCRIPTION OF THE INVENTION

[0012]

　Anti-tumor activity of antibodies against caprin-1 polypeptide for use in the present invention, as described below, in vitro, or show a cytotoxic activity via immune cells against tumor cells expressing the polypeptide by examining whether or can be evaluated by examining the inhibition of tumor growth to tumor-bearing animal in vivo.

[0013]

　Antibodies against the above caprin-1 according to the present invention can be a monoclonal or polyclonal antibody but is preferably a monoclonal antibody, as long as may be any type of antibody capable of exhibiting anti-tumor activity, e.g. , recombinant antibodies (for example, synthetic antibodies, multi-specific antibodies (eg, bispecific antibodies), a humanized antibody, chimeric antibody, single-chain antibody (scFv), etc.), a human antibody, those of antibody fragment (for example, Fab, F (ab ') 2 comprising, Fv, etc.). These antibodies and fragments thereof also can be prepared by methods known to those skilled in the art. Also, if the subject is a human, it is desirable to avoid or suppress rejection reaction is a human antibody or a humanized antibody.

[0014]

　The "caprin-1 protein that specifically binds" specifically binds to caprin-1 protein, it means that does not substantially bind other proteins and.

[0015]

　Further, the subject is a subject of treatment and / or prevention of cancer in the present invention, human, companion animals, livestock, a mammal such as a sport animals, preferably the subject is human.

[0016]

　Hereinafter, production of antigens related to the present invention, the production of antibodies and the pharmaceutical compositions described.

[0017]

　
　protein or fragment thereof is used as the sensitizing antigen for obtaining antibodies against caprin-1 according to the present invention, human, dog, cat, cow, horse, mouse, rat, chicken etc., it is not limited to the animal species from which it is derived. However, it is preferably selected in consideration of their compatibility with the parent cells used for cell fusion. In general, preferably protein derived from mammals, in particular proteins of human origin are preferred. For example, if CAPRIN-1 is of the human CAPRIN-1, it is possible to use a cell or the like that expressed human CAPRIN-1 protein or its partial peptide, human CAPRIN-1.

[0018]

　Nucleotide sequence and amino acid sequence of the human CAPRIN-1 and its homologs are, for example, to access the GenBank (US NCBI), BLAST, an algorithm such as FASTA (Karlin and Altschul, Proc Natl Acad Sci USA, 90:.... 5873 .. -5877,1993; Altschul et al, Nucleic Acids Res 25: 3389-3402, can be obtained by utilizing 1997).

[0019]

　In the present invention, the human caprin-1 nucleotide sequence (SEQ ID NO: 16 or 18) or the amino acid sequence (SEQ ID NO: 17 or 19) as a reference, the nucleotide sequence or amino acid sequence and 70% of these ORF or mature portion 100%, preferably 80% to 100%, more preferably 90% to 100%, more preferably from 95% to 100%, for example, 97% to 100%, 98% and 100%, 99% and 100%, or 99 When a nucleic acid or protein consisting of the sequence having a .5% to 100% sequence identity to compare amino acid sequences of which will caprin-1 target (SEQ ID NO: 17 and SEQ ID NO: 19, differences amino acid residues 690 of subsequent to.). Here, "% sequence identity" of two sequences, when not introduce the to or gap the gap, which is to maximize the degree of similarity (or degree of coincidence) alignment (alignment), amino acids ( or it means a percentage (%) of the total number of bases) (identical amino acids to include a number of gaps) (or base).

[0020]

　Fragment of caprin-1 protein from amino acid long epitope antibodies is the smallest unit that recognizes (antigenic determinant) and has a length less than the full length of the protein. Epitopes, mammals, preferably humans, refers to a polypeptide fragment having antigenicity or immunogenicity, its minimum unit, about 7 to 12 amino acids, for example, composed of 8-11 amino acids.

[0021]

　Above, a polypeptide fragment containing the human caprin-1 protein or its partial peptide, for example, Fmoc method (-fluorenylmethyloxycarbonyl method), tBoc method (t-butyloxycarbonyl method) synthesized by chemical synthesis methods such can be (Japan biochemical Society, ed., biochemistry experimental course 1, chemical IV of protein, chemical modification and peptide synthesis, Tokyo Kagakudojin (Japan), 1981). It can also be synthesized by a conventional method using various commercially available peptide synthesizers. In addition, well-known genetic engineering techniques (Sambrook et al., Molecular Cloning, Second Edition, Current Protocols in Molecular Biology (1989), Cold Spring Harbor Laboratory Press, Ausubel et al., Short Protocols in Molecular Biology, 3rd ed., A compendium of Methods from Current Protocols in Molecular Biology (1995), with John Wiley & Sons, etc.), a polynucleotide encoding the polypeptide is prepared and introduced into a host cell incorporating said polynucleotide into an expression vector, said host by producing a polypeptide in a cell, it is possible to obtain a human caprin-1 proteins and polypeptide fragments thereof of interest.

[0022]

　The polypeptide encoding polynucleotide, by a conventional method using a known genetic engineering technique or a commercially available nucleic acid synthesizer can be readily prepared. For example, DNA comprising the nucleotide sequence of the human caprin-1 gene is prepared by performing PCR using a pair of primers designed to the human chromosomal DNA or cDNA library was used as template, you can amplify the nucleotide sequence can do. The reaction conditions for PCR may be appropriately set, for example, thermostable DNA polymerase (eg, Taq polymerase, Pfu polymerase, etc.) and Mg 2+ using containing PCR buffer, 30 sec at 94 ° C. (denaturation), at 55 ° C. 30 seconds to 1 minute (annealing), and the reaction process consisting of 2 minutes at 72 ° C. (extension) as one cycle, for example, after 30 cycles, there may be mentioned a condition of reacting at 72 ° C.. 7 minutes, not limited thereto. Method for PCR, the conditions and the like, eg, Ausubel et al, Short Protocols in Molecular Biology, 3rd ed., A compendium of Methods from Current Protocols in Molecular Biology (1995), according to John Wiley & Sons (particularly Chapter 15) It is.

[0023]

　Further, based on the nucleotide sequence and caprin-1 protein amino acid sequence information of caprin-1 gene, to prepare a suitable probe or a primer, by screening a cDNA library of human or the like by using it, the desired DNA it can be isolated. cDNA libraries, cells expressing the protein of caprin-1, is preferably produced from an organ or tissue. Examples of such cells or tissues, other testicular, leukemia, breast cancer, lymphoma, brain cancer, lung cancer, pancreatic cancer, colon cancer, renal cancer, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, oesophageal cancer, derived from sarcoma, mast cell tumors, liver cancer, gallbladder cancer, melanoma, adrenocortical cancer, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer or cancer or tumors of the head and neck cancer, etc. it is a cell or tissue. Preparation of the probes or primers, construction of cDNA libraries, screening of cDNA libraries, as well as operations such as cloning of target genes are known to those skilled in the art, for example, Sambrook et al, Molecular Cloning, 2nd edition, the Current Protocols in Molecular Biology (1989), it can be carried out according to the method described in Ausbel et al. (supra) and the like. Thus from the obtained DNA as a, it is possible to obtain DNA encoding human caprin-1 protein or its partial peptide.

[0024]

　As the host cells to which the expression vector is introduced, the long polypeptide a cell capable of expressing may be of any type, such as E. coli as an example of prokaryotic cells, monkey kidney cells as an example of a eukaryotic cell COS1 , mammalian cells such as Chinese hamster ovary cells CHO, human embryonic kidney cell line HEK293, mouse fetal skin cell line NIH3T3, budding yeast, yeast cells, such as fission yeast, silkworm cells, Xenopus egg cell, and the like, to these but it is not limited.

[0025]

　When using a prokaryotic cell as the host cell, as the expression vector, capable of replication origin in prokaryotic cells, a promoter, a ribosome binding site, multiple cloning site, terminator, drug resistance gene, an expression vector having an auxotrophic complementary gene, etc. used. As an expression vector for E. coli, pUC system, pBluescriptII, pET expression system, pGEX expression systems, and the like can be exemplified. Incorporating a DNA encoding the above polypeptide into such an expression vector, after a prokaryotic host cell transformed with the vector, by culturing the obtained transformant, a prokaryotic polypeptide which the DNA encodes it can be expressed in a host cell. In this case, the polypeptide can also be expressed as fusion proteins with other proteins.

[0026]

　When using eukaryotic cells as host cells, as expression vectors, promoter, splice region, an expression vector for eukaryotic cells having a poly (A) addition site and the like is used. Such expression vector, pKA1, pCDM8, pSVK3, pMSG , pSVL, pBK-CMV, pBK-RSV, EBV vector, pRS, pcDNA3, pYES2, and the like can be exemplified. Similar to the above, incorporating a DNA encoding the above polypeptide into such an expression vector, after a eukaryotic host cell with the vector and transformed by culturing the obtained transformant, the DNA encodes the by which polypeptides can be expressed in eukaryotic host cells. PIND / V5-His as an expression vector, when using pFLAG-CMV-2, pEGFP- N1, pEGFP-C1 or the like, His tag (e.g., (the His) . 6 ~ (the His) 10 ), FLAG tag, myc tag, HA tag, as a fusion protein with the addition of GFP and various tags, it is possible to express the said polypeptide.

[0027]

　Introduction into the host cell expression vectors, electroporation, calcium phosphate method, liposome method, DEAE dextran method, microinjection, viral infection, lipofection, can be employed known methods bonds such as a cell membrane permeable peptide.

[0028]

　In order to isolate and purify a polypeptide of interest from host cells can be conducted by combining known separation operations. For example, treatment with denaturing agents or surfactants such as urea, sonication, enzymatic digestion, salting out or solvent fractional precipitation, dialysis, centrifugation, ultrafiltration, gel filtration, SDS-PAGE, isoelectric focusing , ion exchange chromatography, hydrophobic chromatography, affinity chromatography, but reverse phase chromatography, and the like, but are not limited to.

[0029]

　For producing antibodies according to the present invention, the antigen produced as described above can be used as a street sensitizing antigen described later.

[0030]

　
　antibodies (immunoglobulins) are usually heteromultimeric glycoproteins comprising a heavy chain and two light chains of at least two. Apart from IgM, immunoglobulin is two identical light (L) chains and two identical heavy (H) hetero tetrameric glycoproteins of about 150kDa composed of chains. Typically, each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide bonds between the heavy chains of different immunoglobulin isotypes varies. Each heavy and light chain also has intrachain disulfide bonds. Each heavy chain has a variable domain (VH region) at one end, followed by a number of constant regions. Each light chain has a variable domain (VL region), has one constant region at the edge of the opposite. The constant region of the light chain is aligned with the first constant region of the heavy chain and the light chain variable domain is aligned with the variable domain of the heavy chain. The variable domain of an antibody specific regions confer binding specificity to the antibody indicate a particular variability called complementarity determining region (CDR). Moiety that is relatively conserved in the variable region framework regions; are called (framework region FR). Variable domains of complete heavy and light chain comprises three complementarity determining regions are connected by four framework regions structure, i.e., FRl from N-terminal in sequence, CDR1, FR2, CDR2, FR3 , CDR3, and FR4 There consists of a structure consolidated. Three complementarity determining region is CDRH1 in the heavy chain in the order from the N-terminal, CDRH2, CDRH3, are similarly called CDRL1, CDRL2, CDRL3 at light chain. The binding specificity of the antibody to the antigen, CDRH3 is the most important. Also, CDRs of each chain are held together in close proximity by the framework regions, which contributes to the formation of the antigen together with complementarity determining regions from the other chain. The constant region does not contribute directly to the thing to which the antibody binds to the antigen, a variety of effector function, for example, antibody-dependent cellular cytotoxicity involvement in the activity (ADCC), phagocytosis via binding to Fcγ receptor ( ADCP), shows neonatal Fc receptor (half-life / clearance rate via FcRn), complement-dependent cytotoxicity via the C1q component of the complement cascade (CDC).

[0031]

　
　anti caprin-1 antibody in the present invention refers to an antibody having a full length or a fragment and immunological reactivity thereof caprin-1 protein.

[0032]

　Here, "immunological reactivity" is meant an antibody and caprin-1 antigen (CAPRIN-1 full length or its partial polypeptide of the protein) and are binding properties in vivo. Failure tumor cells through such binding to caprin-1 antibodies of the present invention (e.g., killing, inhibition or regression) functions are exhibited. Antibodies of the present invention, the tumor in conjunction with caprin-1 protein, for example, breast cancer, renal cancer, pancreatic cancer, colon cancer (e.g., colon cancer), lung cancer, brain cancer, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mast cell tumors, melanoma, adrenocortical cancer, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer or head and neck parts such as cancer can be failure of the.

[0033]

　Antibodies of the present invention are preferably not particularly limited as long as it is a monoclonal antibody, synthetic antibodies, multispecific antibodies (e.g. diabodies, etc. triabodies), human antibodies, humanized antibodies, chimeric antibodies, single chain antibodies, antibody fragments (e.g., Fab, F (ab ') 2 , Fv) and the like. The antibodies can be of any class of immunoglobulin molecules, e.g., IgG, IgE, IgM, IgA , IgD and IgY or any subclass, for example, an IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 , and the like.

[0034]

　Antibodies further, in addition to glycosylation, deglycosylation, acetylation, formylation, amidation, phosphorylation, or pegs may be modified by (PEG) and the like.

[0035]

　The following illustrates the production of a variety of monoclonal antibody.

[0036]

　For example, immunized by administering the breast cancer cell line SK-BR-3 and the like which express caprin-1 in mice, extracts the spleen from the mouse, on the cell separation, fusing said cell and mouse myeloma cells , from the resulting fused cells (hybridomas), selecting clones which produce antibodies with the cancer cell growth inhibitory action. The monoclonal antibody-producing hybridoma having a cancer cell growth inhibitory action isolated and culturing the hybridoma and purified antibodies by culture supernatant General affinity purification techniques, is possible to prepare antibodies of the present invention is there.

[0037]

　The hybridoma producing the monoclonal antibody can, for example, can be produced also in the following manner. First of all, according to known methods, immunization of animals with a sensitizing antigen. As a general method, the sensitizing antigen is performed by injecting intraperitoneally or subcutaneously in mammals. Specifically, diluting the sensitizing antigen in an appropriate amount of PBS (Phosphate-Buffered Saline), physiological saline or the like, a conventional adjuvant desired to those suspended, for example, Freund's complete adjuvant, after emulsification , administered several times every 4 to 21 days in a mammal. It is also possible to use a suitable carrier at the time of immunization of the sensitizing antigen.

[0038]

　In this way to immunize a mammal, after the desired antibody levels were confirmation of the increase in the serum, the immune cells were harvested from the mammal and subjected to cell fusion, as the preferred immune cells, especially the spleen cells and the like.

[0039]

　As the other parent cell to be fused with the immune cells, using a myeloma cell of a mammal. As the myeloma cells, various known cell lines, for example, P3U1 (P3-X63Ag8U1), P3 (P3x63Ag8.653) (J. Immunol. (1979) 123, 1548-1550), P3x63Ag8U. 1 (Current Topics in Microbiology and Immunology (1978) 81, 1-7), NS-1 (Kohler. G. and Milstein, C. Eur. J. Immunol. (1976) 6, 511-519), MPC-11 (Margulies. D.H. et al., Cell (1976) 8, 405-415), SP2 / 0 (Shulman, M. et al., Nature (1978) 276, 269-270), FO (deSt. Groth , S.F. et al., J. Immunol. Methods (1980) 35, 1-21), S194 (Trowbridge, I.S. J.Exp.Med. (1978) 148, 313-323), R210 ( Galfre, G. et al., Nature (1979) 277, 131-133), such as 240E-1,240E-W and 240E-W2 is preferably used.

[0040]

　The cell fusion of the immune cells and the myeloma cells, a known method is basically, for example, Kohler and the method of Milstein (Kohler, G. and Milstein, C. Methods Enzymol. (1981) 73, 3-46) it can be performed according to equal.

[0041]

　More specifically, the above cell fusion, for example, carried out in the presence of a cell fusion promoter in a conventional nutrient culture medium. Examples of the fusion accelerator, polyethylene glycol (PEG), is used Sendai virus (HVJ) or the like, can be added using an auxiliary agent such as dimethyl sulfoxide in order to enhance efficiency of the fusion optionally.

[0042]

　The ratio of immunocytes to myeloma cells may be set arbitrarily. As the culture medium used for cell fusion, for example, a suitable RPMI1640 culture medium for the growth of the above myeloma cell lines, MEM culture medium, other conventional culture medium used for this type of cell culture can be used, further It may be used in combination with serum supplement such as fetal calf serum (FCS).

[0043]

　In cell fusion, the well mixed with immune cells and the culture solution in a predetermined amount of the myeloma cells, pre-37 ℃ about the warmed PEG solution (for example, having an average molecular weight of 1000-6000) the normal 30-60% was added at a concentration of (w / v), and form the hybridomas of interest by mixing. Subsequently, sequential addition of a suitable culture liquid, it is preferable to remove the cell fusion agents etc. which are undesirable for the growth of the hybridoma by repeating an operation of removing the supernatant by centrifugation.

[0044]

　Hybridomas obtained in this manner, the standard selection medium, for example, is selected by culturing in HAT medium (hypoxanthine, aminopterin, and culture medium containing thymidine). The culture in the HAT culture medium is, cells other than the desired hybridoma (non-fusion cells) for a period of time sufficient to kill (usually, a few days to several weeks). Then, the conventional limiting dilution is performed to screen and single cloning of a hybridoma producing an antibody of interest.

[0045]

　In addition to obtaining the above hybridoma by immunizing non-human animals with antigens, it was sensitized with human lymphocytes, for example, protein infected human lymphocytes to EB virus in in vitro, protein-expressing cells or the lysate-sensitive create lymphocytes myeloma cells having a permanent division potential of human origin, for example, U266 (registration number TIB196) and fused, desired activity (for example, cell growth inhibition activity) is also possible to obtain hybridomas that produce human antibodies having the it can.

[0046]

　Hybridomas producing monoclonal antibodies prepared in this way, it is possible to subcultured in a conventional culture liquid, or can be long-term storage in liquid nitrogen.

[0047]

　In other words, by using the cells expressing the desired antigen or desired antigen as a sensitizing antigen and is immunized in the conventional method of immunization, it is fused with known parent cells obtained immune cells by a conventional cell fusion method by conventional screening methods, a monoclonal antibody-producing cells (hybridomas) can be prepared by screening.

[0048]

　Preparation of antigens, for example, a method using animal cells method using (Kohyo 2007-530068) and baculovirus (e.g., International Publication No. W098 / 46 777 No., etc.) can be performed according to such. If the antigen has low immunogenicity, conjugated with macromolecules having immunogenic such as albumin, it may be carried out immunological. The antigen may be immunized by administration with an adjuvant.

[0049]

　Antibodies of the present invention, furthermore, cloning the antibody gene from the hybridoma, incorporating in a suitable vector, which is introduced into a host, obtained as recombinant antibodies generated with genetic engineering techniques it is possible (for example, Carl, reference A.K. Borrebaeck, James, W. Larrick, THERAPEUTIC MONOCLONAL ANTIBODIES, Published in the United Kingdom by MACMILLAN PUBLISHERS LTD, 1990). Specifically, cDNA is synthesized in the variable region (V region) of the antibody using reverse transcriptase from the hybridoma mRNA. If Rarere obtained DNA encoding the V region of the antibody of interest, which was linked to the DNA encoding the desired antibody constant region (C region), which is then integrated into an expression vector. Alternatively, the DNA encoding the V region of the antibody may be integrated into an expression vector containing the DNA encoding the C region of the antibody. Expression control region, for example, incorporated into the expression vector so as to be an expression enhancer, under the control of the promoter. Subsequently, the expression vector transformed into a host cell and the antibody can be expressed.

[0050]

　Anti-caprin-1 antibody of the present invention preferably is characterized in that it is a monoclonal antibody, the monoclonal antibodies are human monoclonal antibodies, non-human animal monoclonal antibodies (e.g., murine monoclonal antibodies, rat monoclonal antibodies, rabbit monoclonal antibodies, chicken monoclonal antibody, etc.), include chimeric monoclonal antibodies, and the like. Monoclonal antibodies, non-human mammal immunized with caprin-1 protein or a fragment thereof (e.g., mouse, human antibody-producing mouse, chicken, rabbit, etc.) culturing a hybridoma obtained by fusion of spleen cells and myeloma cells from It can be made by. Chimeric antibodies are antibodies prepared by combining sequences derived from different animal, for example, the heavy chain of murine antibody, the heavy chain variable region and a human antibody light chain, is an antibody or the like consisting of the constant region of the light chain . Preparation of the chimeric antibody can be carried out using known methods, for example, to connect the DNA encoding the DNA and human antibody C region encoding an antibody V region, and introduced into a host by incorporating this into an expression vector to produce the antibody.

[0051]

　In the embodiment described below, a plurality of humanized monoclonal antibody and human - rabbit chimeric monoclonal antibodies are prepared, strong anti-tumor effect was confirmed. These monoclonal antibodies are all represented by the amino acid sequence of the heavy chain variable region CDR1 represented by the amino acid sequence of SEQ ID NO: 1 to (VH region) is represented by the amino acid sequence of SEQ ID NO: 2 CDR2 and SEQ ID NO: 3 wherein the CDR3, the light chain variable region (VL region), CDRl represented by the amino acid sequence of SEQ ID NO: 4, a CDR3 represented by the amino acid sequence of CDR2 and SEQ ID NO: 6 shown by the amino acid sequence of SEQ ID NO: 5 including. Each of these monoclonal antibodies, of SEQ ID NO: 7 of the VH region and SEQ ID NO: 11 having an amino acid sequence humanized antibody # 0 consisting of the VL region having an amino acid sequence, the VH region and SEQ ID NO: 11 having an amino acid sequence of SEQ ID NO: 8 humanized antibody # 2 consisting of the VL region having an amino acid sequence of the VH region and SEQ ID NO: 12 having an amino acid sequence of the humanized antibody # 1, SEQ ID NO: 8 consisting of the VL region having an amino acid sequence, the amino acid sequence of SEQ ID NO: 8 humanized antibody comprising the VL region having an amino acid sequence of the VH region and SEQ ID NO: 13 having # 3, humanized antibodies # 4 consisting of the VL region having an amino acid sequence of the VH region and SEQ ID NO: 12 having an amino acid sequence of SEQ ID NO: 7 , VL having the amino acid sequence of the VH region and SEQ ID NO: 15 having a humanized antibody # 5, the amino acid sequence of SEQ ID NO: 7 consisting of the VL region having an amino acid sequence of the VH region and SEQ ID NO: 13 having an amino acid sequence of SEQ ID NO: 7 sequence as VH region having a humanized antibody # 7, the amino acid sequence of SEQ ID NO: 9 consisting of the VL region having an amino acid sequence of the VH region and SEQ ID NO: 15 having an amino acid sequence of the humanized antibody # 6, SEQ ID NO: 8 consisting of region humanized antibody # 8 consisting of the VL region having an amino acid sequence of ID NO: 15, SEQ ID NO: 10 humanized antibody # 9 consisting of the VL region having an amino acid sequence of the VH region and SEQ ID NO: 14 having an amino acid sequence of SEQ ID NO: 10 human consisting of the VL region having an amino acid sequence of the VH region and SEQ ID NO: 21 having an amino acid sequence of the humanized antibody # 10, and SEQ ID NO: 20 consisting of the VL region having an amino acid sequence of the VH region and SEQ ID NO: 15 having an amino acid sequence - including a rabbit chimeric antibody.

[0052]

　Humanized antibodies are referred to as modified antibody with reconstruction (reshaped) human antibody. Humanized antibodies, complementarity determining regions of an antibody derived from an immunized animal, is constructed by grafting the complementarity determining region of a human antibody. The general gene recombination techniques are also known.

[0053]

　More specifically, for example, made to have a mouse antibody, a DNA sequence which was designed to connect the framework regions of complementarity determining region of a human antibody of rabbit antibody and chicken antibody, sections overlapping at the ends synthesized by PCR from several oligonucleotides. The resulting DNA was ligated with DNA encoding human antibody constant region, and then is integrated into an expression vector, which is obtained by producing introduced into a host (European Patent Application Publication No. EP 239 400, WO WO96 / reference No. 02576). Framework region of a human antibody, which is connected via a complementarity determining region, the complementarity determining region that forms a favorable antigen-binding site is selected. If necessary, the complementarity determining region of reshaped human antibody may be replaced amino acids in the framework region in the variable region of the antibody so as to form an appropriate antigen binding site (Sato K. et al., Cancer Research 1993, 53: 851-856). Also, it may be substituted in the framework regions from various human antibodies (see International Publication No. WO99 / ​​51743).

[0054]

　In making chimeric antibodies and humanized antibodies, variable region (for example, FR) and the amino acid in the constant region may be substituted with other amino acids.

[0055]

　Amino acid substitutions, one or a plurality, for example, less than 15, less than 10, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or more than two amino acids, preferably a substituted 1 to 9 amino acids , and the substituted antibody should be functionally equivalent to the unsubstituted antibody.

[0056]

　Herein, "functionally equivalent", similar biological or biochemical activity as the antibody of the antibody present invention of interest, it is specifically has a function of damaging tumors, when applied to human It refers to such that essentially cause the rejection. Such activities include, for example, it can be exemplified cytostatic activity or binding activity.

[0057]

　For the preparation of a a polypeptide functionally equivalent polypeptides, as the methods well known to those skilled in the art, a method of amino acid substitutions by introducing mutations into polypeptides are known. For example, those skilled in the art, site-specific mutagenesis (Hashimoto-Gotoh, T. et al., (1995) Gene 152, 271-275, Zoller, MJ., And Smith, M. (1983) Methods Enzymol . 100, 468-500, Kramer, W. et al., (1984) Nucleic Acids Res. 12, 9441-9456, Kramer, W. and Fritz, HJ., (1987) Methods Enzymol. 154, 350-367, Kunkel, TA., (1985) Proc. Natl. Acad. Sci. USA. 82, 488-492, Kunkel (1988) Methods Enzymol. 85, using 2763-2766) and the like, optionally amino acid substitutions in an antibody of the present invention by introducing, can be prepared functionally equivalent antibody antibody.

[0058]

　If you want to introduce the amino acid substitution, the substitution, it is desirable conservative amino acid substitution. A conservative amino acid substitution, charge, side chain, polarity, is a substitution between amino acid that is similar in nature, such as aromatic. The similar amino acid nature, for example, basic amino acids (arginine, lysine, histidine), acidic amino acids (aspartic acid, glutamic acid), uncharged polar amino acids (glycine, asparagine, glutamine, serine, threonine, cysteine, tyrosine), apolar sexual amino acids (leucine, isoleucine, alanine, valine, proline, phenylalanine, tryptophan, methionine), branched chain amino acids (leucine, valine, isoleucine), aromatic amino acids can be classified into (phenylalanine, tyrosine, tryptophan, histidine) and the like.

[0059]

　The modified antibody can include, for example, an antibody bound to various molecules such as polyethylene glycol (PEG). In the modified antibodies of the present invention, the substance to be coupled is not limited. To obtain such modified antibodies can be obtained by the obtained antibody chemically modified. These methods have already been established in this field.

[0060]

　Caprin-1 protein or antibody that recognizes caprin-1 fragment polypeptide comprising the same, can be obtained by methods known to those skilled in the art. For example, as determined by epitope conventional methods of anti-caprin-1 antibody that recognizes caprin-1 protein (e.g., a method in the identification of epitopes for epitope mapping and below), poly having an amino acid sequence contained in the epitope a method of producing antibodies the peptide as an immunogen, to determine the epitope of the produced antibodies by conventional methods, can be anti-caprin-1 antibody and epitope obtained by a method such as to select the same antibody.

[0061]

　Antibodies of the present invention, the antibody with caprin-1 and immunological reactivity, caprin-1 antibody that specifically recognizes, or caprin-1 and an antibody that specifically binds, cytotoxic activity against cancer , or an antibody that shows a tumor growth inhibitory effect. The antibody is preferably rejection in a subject animal is an antibody having a structure as little or no avoidance of administering it. Such antibodies, for example, when the target animal is a human, a human antibody, a humanized antibody, a chimeric antibody (e.g., a human - rabbit chimeric antibodies), single chain antibodies, bispecific antibodies and the like. These antibodies, or the variable region of the heavy and light chains are of human origin antibodies, complementarity determining regions of the variable regions of the heavy and light chains from a non-human animal antibody (CDR1, CDR2 and CDR3) or those consisting of a framework region from a human antibody (FR1, FR2, FR3 and FR4), or are those variable regions of the heavy and light chains from a non-human animal antibody, and the heavy and light the constant region of the chain is a recombinant antibody is derived from a human antibody. Preferred antibodies are the previous two antibodies.

[0062]

　These recombinant antibodies can be produced as follows. Monoclonal antibodies from antibody-producing cells such as hybridomas against human CAPRIN-1 (e.g., human monoclonal antibodies, murine monoclonal antibodies, rat monoclonal antibodies, rabbit monoclonal antibodies, chicken monoclonal antibody, etc.) to clone DNA encoding, which was a template DNA encoding the light and heavy chain variable regions of the antibodies produced by an RT-PCR method or the like on, for example, Kabat EU numbering system (Kabat et al, Sequences of Proteins of Immunological Interest, 5thEd. Public Health Service, National Institute of Health, Bethesda, Md. (1991) of the light and heavy chains based on), the sequence of each of the variable region, or sequence of each CDR1, CDR2, CDR3, or sequence of each FR1, FR2, FR3, FR4 it can be determined.

[0063]

　Furthermore, the DNA encoding the DNA or the complementarity determining regions coding for these respective variable regions of a gene recombination technique (Sambrook et al, Molecular Cloning A Laboratory Manual, Cold Spring Harbor Laboratory Press (1989)) or a DNA synthesizer It produced using. Here, the human monoclonal antibody-producing hybridomas, human antibody-producing animal (e.g., a mouse) After immunizing human caprin-1 to, be produced by fusing the spleen cells with myeloma cells excised from the immunized animal can. Apart from this, if necessary, to produce DNA encoding the variable regions and constant regions of the light chain or the heavy chain derived from a human antibody using genetic engineering technique or a DNA synthesizer.

[0064]

　In the case of humanized antibody, a CDR coding sequences in the DNA encoding the variable region of the light chain or the heavy chain derived from a human antibody, their corresponding non-human animal (e.g., mouse, rat, rabbit, chicken by substituting CDR coding sequences from an antibody etc.), it is possible to produce a DNA encoding the humanized antibody. For example, if the CDR coding sequences derived from human antibody of the mouse antibody from the CDR coding sequence and substituted humanized antibody, the variable region is a human FR1 from the N-terminal side, mouse CDR1, human FR2, mouse CDR2, human FR3, mouse CDR4, and a in the order of human FR4.

[0065]

　In the case of the chimeric antibody, a non-human animal (eg, mouse, rat, rabbit, chicken, etc.) each of the DNA encoding the variable region of the light chain or the heavy chain of the antibody derived from the light chain or heavy derived from a human antibody by ligated to DNA encoding the constant region of the chain, it is possible to produce a DNA encoding the chimeric antibody.

[0066]

　In the case of single-chain antibodies, the antibody is an antibody linked linearly through a linker and a heavy chain variable region and light chain variable region, the coding DNA, a linker encoding the heavy chain variable region DNA , and it is possible to create a DNA encoding the single chain antibody by binding the DNA encoding the light chain variable region. Here, both a heavy chain variable region and light chain variable region, or is derived from a human antibody, or, non-human only the complementarity determining regions animals (e.g., mice, rats, rabbits, chickens, etc.) derived from are those of a human antibody-derived substituted by complementarity determining region of an antibody. Further, the linker is made ​​12-19 amino acids, e.g., of 15 amino acids (G 4 S) 3 (G. -B Kim et al., Protein Engineering Design and Selection 2007, 20 (9):. 425-432) include It is.

[0067]

　Bispecific antibodies (e.g., diabody) in the case of the antibody are two different epitopes specifically capable of binding antibodies, for example, DNA encoding heavy chain variable region A, the light chain variable region B DNA encoding, DNA encoding the heavy chain variable region B, and binds the DNA encoding the light chain variable region a in this order (however, the DNA and heavy chain variable region B which encodes a light chain variable region B the encoding DNA is attached via DNA encoding a linker as described above.) can be prepared DNA encoding the bispecific antibody by. Here, both a heavy chain variable region and light chain variable region, or is derived from a human antibody, or, non-human only the complementarity determining regions animals (e.g., mice, rats, rabbits, chickens, etc.) derived from are those of a human antibody-derived substituted by complementarity determining region of an antibody.

[0068]

　The recombinant DNA is prepared as described above, incorporated into one or more suitable vectors, which were introduced into a host cell (e.g., mammalian cells, yeast cells, insect cells, etc.), (co) expression is to be able to produce a recombinant antibody by (P.J. Delves, aNTIBODY PRODUCTION ESSENTIAL TECHNIQUES, 1997 WILEY, P Shepherd and C. Dean, Monoclonal antibodies, 2000 OXFORD UNIVERSITY PRESS;..... J. W. Goding, Monoclonal Antibodies:.. principles and practice, 1993 ACADEMIC PRESS).

[0069]

　The antibody of the present invention manufactured by the above method, for example, a light chain variable region comprising the heavy chain variable region SEQ ID NO: 4, 5 and 6 comprising SEQ ID NO: 1, 2 and 3 obtained in the Examples below the following antibodies comprising bets (a) ~ (l), and the like.

[0070]

　(A) a heavy chain variable region and light chain variable region composed of the antibody of SEQ ID NO: 15 SEQ ID NO: 8
　heavy chain variable region and an antibody composed of light chain variable region of SEQ ID NO: 15 (b) SEQ ID NO: 10
　(c) a heavy chain variable region and light chain variable region composed of the antibody of SEQ ID NO: 15 SEQ ID NO: 7
　antibody composed of light chain variable region of the heavy chain variable region and SEQ ID NO: 13 (d) SEQ ID NO: 8
　(e) a heavy chain variable region and light chain variable region composed of the antibody of SEQ ID NO: 12 SEQ ID NO: 7
　a heavy chain variable region and an antibody composed of light chain variable region of SEQ ID NO: 12 (f) SEQ ID NO: 8
　(g) a heavy chain variable region and light chain variable region composed of the antibody of SEQ ID NO: 13 SEQ ID NO: 7
　antibody composed of a heavy chain variable region and light chain variable region of SEQ ID NO: 14 of (h) SEQ ID NO: 10
　(i) a heavy chain variable region and light chain variable region composed of the antibody of SEQ ID NO: 11 SEQ ID NO: 8
　antibody composed of light chain variable region of the heavy chain variable region and SEQ ID NO: 11 (j) SEQ ID NO: 7
　(k) a heavy chain variable region and an antibody composed of light chain variable region of SEQ ID NO: 15 SEQ ID NO. 9
　(l) an antibody composed of light chain variable region of the heavy chain variable region and SEQ ID NO: 21 SEQ ID NO: 20
　here, each of the amino acid sequence shown in SEQ ID NO: 1, 2 and 3, a CDR1, CDR2 and CDR3 of the heavy chain variable regions of rabbit antibody, respectively the amino acid sequence shown in SEQ ID NO: 4, 5 and 6, the rabbit antibodies a CDR1, CDR2 and CDR3 of the light chain variable region.

[0071]

　Furthermore, humanized antibodies of the present invention, chimeric, single chain or bispecific antibodies, for example, the following antibodies (i) ~ (xiv).

[0072]

　(I) the variable region of the heavy chain comprises an amino acid sequence or derivatives thereof of the framework region amino acid sequences and derived from a human antibody SEQ ID NO: 1, 2 and 3, and the variable region of the light chain is SEQ ID NO: 4, 5 and 6 amino acid sequences and human antibody-derived framework region amino acid sequence or antibodies comprising the substitution product of.

[0073]

　(Ii) the variable region of the heavy chain comprises an amino acid sequence or derivatives thereof of the framework region amino acid sequences and derived from a human antibody SEQ ID NO: 1, 2 and 3, and the constant region of the heavy chain from a human antibody amino acid comprises a sequence, and comprises an amino acid sequence or derivatives thereof of the framework region amino acid sequences and derived from a human antibody variable region of the light chain is SEQ ID NO: 4, 5 and 6, and the constant region of the light chain is a human antibody antibody comprising an amino acid sequence derived from.

[0074]

　(Iii) comprises an amino acid sequence of the variable region of the heavy chain SEQ ID NO: 8, and the constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 15 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0075]

　(Iv) comprises an amino acid sequence of the variable region of the heavy chain SEQ ID NO: 10, and constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 15 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0076]

　(V) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 7, and the constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 15 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0077]

　(Vi) comprises the amino acid sequence of the variable region of the heavy chain SEQ ID NO: 8, and the constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 13 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0078]

　(Vii) comprises the variable region of the heavy chain amino acid sequence of SEQ ID NO: 7, and the constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 12 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0079]

　(Viii) comprises the amino acid sequence of the variable region of the heavy chain SEQ ID NO: 8, and the constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 12 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0080]

　(Ix) wherein the variable region of the heavy chain amino acid sequence of SEQ ID NO: 7, and the constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 13 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0081]

　(X) comprises the amino acid sequence of the variable region of the heavy chain SEQ ID NO: 10, and constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 14 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0082]

　(Xi) comprising the amino acid sequence of the variable region of the heavy chain SEQ ID NO: 8, and the constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 11 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0083]

　(Xii) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 7, and the constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 11 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0084]

　(Xiii) comprises the amino acid sequence of the variable region of the heavy chain SEQ ID NO: 9, and the constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 15 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0085]

　(Xiv) comprises the amino acid sequence of the variable region of the heavy chain SEQ ID NO: 20, and constant region of the heavy chain comprises an amino acid sequence derived from human antibody, and the amino acid sequence of the variable region of the light chain is SEQ ID NO: 21 It includes, and an antibody constant region of the light chain comprising the amino acid sequence derived from a human antibody.

[0086]

　Incidentally, the sequence of the framework regions of the constant region and the variable region of a human antibody heavy and light chains, for example, NCBI: is available (U.S. GenBank, UniGene, etc.) from, for example, for the human IgG1 heavy chain constant region registration number J00228, registration for the human IgG2 heavy chain constant region number J00230, registration for the human IgG3 heavy chain constant region number X03604, registration number for the human IgG4 heavy chain constant region K01316, registration number for the human light chain κ constant region V00557, X64135, X64133 and the like, for the human light chain λ constant region can refer to the sequence such as registration number X64132, X64134.

[0087]

　The antibody preferably has cytotoxic activity and thereby can exert an anti-tumor effect (or anti-tumor activity).

[0088]

　In addition the antibody, as long as it has a specificity that specifically recognize the CAPRIN-1, the sequence of the complementarity determining region of each antibody, in the sequence and / or sequence of the constant region of the framework regions, one or several of substitution of an amino acid, there may be a deletion or addition. Here, the several, preferably means one to nine.

[0089]

　Antibodies of the present invention, the affinity constant Ka for caprin-1 protein or a fragment thereof (k on / k off ) is preferably at least of at least 5 × 10 . 8 M -1 , at least 10 . 9 M -1 , at least 5 × 10 9 M -1 , at least 10 10 M -1 , at least × 5 10 10 M -1 , at least 10 11 M -1 , at least × 5 10 11 M -1 , at least 10 12 M -1 , at least 10 13 M -1 , or at least 10 14 M -1 it is.

[0090]

　One of the mechanisms of anti-tumor effect against caprin-1-expressing cancer cells with an antibody of the present invention, there is an effector cell antibody dependent cellular cytotoxicity of caprin-1 expressing cells (ADCC). Amino acid 1 or several substituents of the heavy chain constant region of an antibody of the present invention, or, the removal of fucose is bound to N- acetylglucosamine in the heavy chain that binds to the constant region N- glycoside-linked sugar chain Accordingly, it is possible to enhance the anti-tumor activity by ADCC of the antibodies of the present invention. Further, the by combining amino acid substitutions and fucose removal of the heavy chain constant region, it is possible to further enhance the anti-tumor activity by ADCC of the antibodies of the present invention.

[0091]

　Further, the antibody was removed fucose attached to N- acetylglucosamine in the heavy chain constant region to bind N- glycoside-linked sugar chain in the present invention may be a alone, also fucose is bound it may be a composition of an antibody that. In the composition of the antibody, it is preferable antibody removal of the fucose is a main component.

[0092]

　Antibody that has been one or several substituting amino acids of the heavy chain constant region, for example, in International Publication No. WO2004 / 063351 Patent, International Publication No. WO2011 / 120135, US Pat. No. 8,388,955, International Publication No. WO2011 / 005481, US Patent 6,737,056 No., it can be prepared by referring to WO WO2005 / 063351. N- glycoside-linked sugar antibody or its production cell fucose has been removed which is added to the N- acetyl glucosamine in the chain in the heavy chain constant region, for example, in US Pat. No. 6,602,684, European Patent No. 1,914,244, US Pat. No. 7,579,170 it can be prepared with reference to. Composition or the cells producing antibodies antibody and fucose removal of the fucose attached is attached to the N- acetyl glucosamine in the heavy chain constant region to join N- glycoside-linked sugar chain, for example, in US patent 8,642,292 it can be prepared by referring to the items.

[0093]

　Antibodies of the present invention can be anti-tumor agent conjugated. The binding of the antibody to the anti-tumor agent, an amino group, a carboxyl group, a hydroxy group, a reactive group with a thiol group and the like (e.g., succinimidyl group, formyl group, 2-pyridyldithio group, Mareiimijiru group, an alkoxycarbonyl group it can be carried out via a spacer having a hydroxy group).

[0094]

　Examples of anti-tumor agents, known anti-tumor agents of the following in the literature, etc., ie, paclitaxel, doxorubicin, daunorubicin, cyclophosphamide, methotrexate, 5-fluorouracil, thiotepa, busulfan, improsulfan, piposulfan, Benzodopa (benzodopa ), carboquone, Metsuredopa (meturedopa), Uredopa (uredopa), alto rate amine (altretamine), triethylene melamine, triethylenephosphoramide, triethylene thiophosphate Hora bromide (triethilenethiophosphoramide), trimethylol Loro melamine (trimethylolomelamine), Buratashin, Buratashinon , camptothecin, bryostatin, kallistatin (callystatin), cryptophycin 1, cryptophycin 8, dolastatin,'s Okaru clarithromycin, eleutherobin, punk Lachi statin, Sarkozy Kuching (sarcodictyin), sponge statin, chlorambucil, Kuroronafajin (chloRNAphazine), Korohosufamido (cholophosphamide), estramustine, ifosfamide, mechlorethamine, black Letter Min-oxide hydrochloride, melphalan, Nobenbichin (novembichin), phenesterine (phenesterine), prednimustine (prednimustine), trofosfamide (trofosfamide), uracil mustard, carmustine, chlorozotocin (chlorozotocin), fotemustine (fotemustine), lomustine, nimustine, ranimustine, calicheamicin (calicheamicin), Dainemaishin, clodronate, esperamicin, aclacinomycin, actinomycin, Orth La clarithromycin (authramycin), azaserine, bleomycin, around Nomaishin (cactinomycin), Karabishin (carabicin), carminomycin, Karujinofirin (carzinophilin), chromomycin, dactinomycin, Detorubishin (detorbicin), 6- diazo-5-oxo -L- norleucine, adriamycin (adriamycin), epirubicin, Esorubishin, idarubicin, Ma Cerro clarithromycin (marcellomycin), mitomycin C, mycophenolic acid (mycophenolic acid), Nogaramaishin (nogalamycin), Oribo clarithromycin (olivomycins), peplomycin, Pot Philo clarithromycin (potfiromycin), puromycin, Keramaishin (quelamycin ), Rodorubishin (rodorubicin), streptonigrin, streptozocin, tubercidin (tubercidin), ubenimex, zinostatin (zinostatin), zorubicin (zorubicin), Denoputerin (denopterin), pteropterin (pteropterin), trimetrexate gravel Sète (trimetrexate), fludarabine ( fludarabine), 6- mercaptopurine, Chiamipurin, thioguanine, ancitabine, azacytidine (azacitidine), 6- Azaurijin (azauridine), carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine (enocitabine), floxuridine (floxuridine); androgens, for example, calusterone (calusterone), Doromosutanoron propionic acid, epitiostanol, mepitiostane, test lactone (testolactone), aminoglutethimide, mitotane, trilostane, Florin acid (frolinic acid), aceglatone, Aldo cyclophosphamide glycoside, aminolevulinic acid, eniluracil , amsacrine (amsacrine), Best Love sill (bestrabucil), Bisantoren (bisantrene), Edatorakiseto (edatraxate), Defofamin (defofamine), Demekorushin (demecolcine), Jiajikon (diaziquone), Eruforunichin (elfornithine), acetic acid Eripuchiniumu (elliptinium ), epothilones (epothilone), Etogurushido (etoglucid), lentinan, lonidamine (lonidamine), maytansine (maytansine), ansamitocin (ansamitocine), mitoguazone (mitoguazone), mitoxantrone, Mopidanmoru (mopidanmol), Nitoraerin (nitraerine), pentostatin , Fenametto (phenamet), pirarubicin, losoxantrone (losoxantrone), podophyllin acid (podophyllinic acid), 2- ethyl hydrazide, procarbazine, Razokisan (razoxane), rhizoxin, schizophyllan, spiro germanium (spirogermanium), Tenyuazon acid (tenuazonic acid), Toriajikon (triaziquone), Roridin (roridine) A, Anguijin (anguidine), urethane, vindesine, dacarbazine, mannomustine (mannomustine), mitobronitol, mitolactol (mitolactol), pipobroman (pipobroman), Gashitoshin (gacytosine), doxetaxel, chlorambucil, gemcitabine (gemcitabine ), 6-thioguanine, mercaptopurine, cisplatin, oxaliplatin, carboplatin, vinblastine, etoposide, ifosfamide, mitoxantrone, vincristine, vinorelbine, Novantrone (novantrone), teniposide, edatrexate (edatrexate), daunomycin, aminopterin, Kiseroda (xeloda ), including ibandronate (ibandronate), irinotecan, topoisomerase inhibitors, difluoro methylol two Chin (DMFO), retinoic acid, capecitabine (capecitabine), as well as their pharmaceutically acceptable salts or derivatives.

[0095]

　Antibody, in the case of anti-tumor agent conjugated to the antibody, as a method of assessing whether exert anti-tumor activity, for example, if the mouse-derived anti-caprin-1 antibody, which binds to the mouse antibody two and the reacted at the same time those with the drug to the next antibody, an anti-tumor effect against human cancer cells can be evaluated in vitro. For example, it is evaluated using saporin anti-human IgG antibody as the second immunotoxin (saporin) was coupled (Hum-ZAP (Advanced Targeting Systems)).

[0096]

　In addition, it is possible to obtain the antibodies of the present invention, by co-administering an anti-tumor agent, a higher therapeutic effect. This technique, for cancer patients CAPRIN-1 is expressed, can be adapted in both before and after surgery. Particularly after surgery, for cancer caprin-1 which has been treated with a conventional anti-tumor agent alone is expressed, extend higher cancer recurrence prevention and survival are obtained.

[0097]

　Antitumor agents used in combined administration with the antibodies of the present invention, for example, can utilize the anti-tumor agent. In particular, cyclophosphamide, paclitaxel, doxetaxel, vinorelbine is preferably used.

[0098]

　Alternatively, the antibodies of the present invention, known in the literature or the like, 211 At, 131 the I, 125 the I, 90 Y-, 186 Re, 188 Re, 153 SM, 212 Bi, 32 P, 175 Lu, 176 Lu, 89 Sr , 64 Cu, 111 it is also possible to combine a radioactive isotope such as in (Hideo Saji, YAKUGAKU ZASSHI 128 (3) 323-332 8 (2008), Jpn). Radioactive isotopes, valid for tumor treatment and diagnosis is desirable. Such radioisotopes are also included in the antitumor agent of the present invention.

[0099]

　
　Antitumor effect against caprin-1-expressing cancer cells by anti-caprin-1 antibody used in the present invention is believed to occur by the following mechanisms such as: effector cell antibodies of the aforementioned caprin-1 expressing cells dependent cellular cytotoxicity (ADCC), and CAPRIN-1 expression antibody-dependent cellular phagocytosis of the cells (ADCP). However, to limit the scope of the present invention by this mechanism is not intended.

[0100]

　Thus, the activity evaluation of anti-caprin-1 antibody used in the present invention, as specifically shown in the following examples, in vitro, said ADCC activity or ADCP activity against cancer cells expressing caprin-1 it can be evaluated by measuring the.

[0101]

　Anti-caprin-1 antibody used in the present invention bind with caprin-1 protein on a cancer cell, by the activity or the like, since it exhibits antitumor activity, is believed to be useful in the treatment or prevention of cancer. That is, the present invention is an active ingredient of anti-caprin-1 antibodies, provides a pharmaceutical composition for the treatment and / or prevention of cancer. When used for the purpose of administering an anti-caprin-1 antibody to the human body (antibody treatment), in order to reduce immunogenicity, it is preferable that a human antibody or a humanized antibody.

[0102]

　In addition, the higher the binding affinity of the CAPRIN-1 protein on the anti-CAPRIN-1 antibody and the surface of cancer cells, with anti-CAPRIN-1 antibody, anti-tumor activity is obtained stronger. Accordingly, the antibodies of the present invention has a caprin-1 protein with high binding affinity, can be expected a stronger anti-tumor effect, it can be adapted as a pharmaceutical composition intended for the treatment and / or prevention of cancer become. Antibodies of the present invention, as the high binding affinity, as described above, the binding constant (affinity constant) Ka (k on / k off ) is preferably at least 5 of 5 × 10 . 8 M -1 , at least 10 . 9 M - 1 , at least × 10 5 9 M -1 , at least 10 10 M -1 , at least × 5 10 10 M -1 , at least 10 11 M -1 , at least × 5 10 11 M -1 , at least 10 12 M -1 , at least 10 13 M -1 , or at least 10 14 M -1 it is preferable to have a.

[0103]

　
　ability of the antibody to bind to caprin-1, such as described in Example, eg, ELISA, Western blotting, binding assay using immunofluorescence and flow cytometry analysis, etc. it can be specified using.

[0104]

　
　antibodies that recognize caprin-1 can be used in immunohistochemistry by methods well known to those skilled in the art. For example, tissue or obtained from the patient during surgery, spontaneously or tissue obtained transfection after xenograft tissue inoculated with cell lines expressing caprin-1 from an animal bearing, paraformaldehyde or acetone fixed frozen sections obtained on, or using a fixed tissue sections and paraffin-embedded in paraformaldehyde, can be tested for reactivity with cAPRIN-1.

[0105]

　For immunohistochemical staining, the antibody reactive against CAPRIN-1, can be stained in a variety of ways. For example, horseradish peroxidase conjugated goat anti-mouse antibody, by reacting the goat anti-rabbit antibody and goat anti-chicken antibody, can be visualized.

[0106]

　
　target of a pharmaceutical composition for the treatment and / or prevention of cancer of the present invention, particularly if the cancer (cells) expressing caprin-1 gene but it is not limited.

[0107]

　The term "tumor" and "cancer" as used herein, refers to a malignant neoplasm are used interchangeably.

[0108]

　The cancer of interest in the present invention, may be any cancer if the cancer expressing CAPRIN-1 protein on the cell membrane surface. Preferably, breast cancer described above, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, obesity astrocytoma, melanoma, adrenocortical cancer, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer or head and neck cancer.

[0109]

　The cancer is, more specifically, for example, breast cancer, complex type mammary gland cancer, mammary gland malignant mixed tumor, intraductal papillary adenocarcinoma, lung adenocarcinoma, squamous cell carcinoma, small cell carcinoma, large cell carcinoma, neuroepithelial glioma is tissue tumors, glioblastoma, neuroblastoma, ependymoma, neuronal tumor, fetal form of neuroectodermal tumors, schwannoma, neurofibroma, meningioma, chronic form lymphocytic leukemia, gastrointestinal lymphoma, gastrointestinal lymphoma, small to medium-cell lymphoma, cecum cancer, ascending colon cancer, descending colon cancer, transverse colon cancer, S-shaped colon cancer, colorectal cancer, ovarian epithelial cancer, embryo cell tumor, stromal cell tumor, pancreatic duct cancer, invasive ductal cancer, pancreatic cancer adenocarcinoma, acinar cell carcinoma, adenocarcinoma squamous cell carcinoma, giant cell tumor, intraductal papillary mucinous tumor, mucinous cyst adenocarcinoma, pancreatic neuroblastoma, head of the pancreas tumors, Frants tumor, serous cystadenocarcinoma, solid papillary carcinoma, gastrinoma, glucagonoma, insulinoma, multiple endocrine neoplasia 1 (Wermer syndrome), non-functional island cell tumors, somatostatinoma, VIP producing tumors, cervical cancer, endometrial cancer, fibrosarcoma, bone and joint meat species, Ewing's sarcoma, Wilms tumor, hepatoblastoma, soft tissue sarcoma, acute leukemia, chronic leukemia, spinal cord tumor, malignant tumor soft, teratoma group tumor , hypopharynx cancer in the head and neck cancer, oropharyngeal cancer, tongue cancer, nasopharyngeal cancer, oral cancer, lip cancer, nasal sinus cancer, encompasses the larynx cancer and the like, but is not limited to these.

[0110]

　Also preferred subject of interest (patient) is a mammal, e.g., primates, pet animals, livestock, a mammal, including a sports animals such as, in particular humans, dogs and cats preferred.

[0111]

　In the case of using the antibodies used in the present invention as a pharmaceutical composition can be formulated by methods known to those skilled in the art. For example, a sterile solution with water or other pharmaceutically acceptable liquid, or in the form of suspensions of injections can be used parenterally. For example, carrier or vehicle pharmacologically acceptable, in particular, sterile water, physiological saline, vegetable oils, emulsifiers, suspending agents, surfactants, stabilizers, flavoring agents, excipients, vehicles, preservatives , in appropriate combination with binders like, it is contemplated be formulated by mixing in a unit dosage form required for pharmaceutical practice generally accepted. The amount of active ingredients in these preparations are intended to ensure that an appropriate dose within the specified range is obtained.

[0112]

　Sterile compositions for injection can be formulated following normal drug implementations using vehicles such as distilled water for injection.

[0113]

　The aqueous solution for injection, for example, physiological saline, an isotonic solution containing glucose and other auxiliary agents, for example, D- sorbitol, D- mannose, D- mannitol, sodium chloride, and the like, with an appropriate dissolution aid , for example, alcohols, in particular ethanol, polyalcohol such as propylene glycol, polyethylene glycol, nonionic surfactants, e.g., polysorbate 80 (TM), may be used in combination with HCO-60.

[0114]

　Sesame oil as the oily liquid, soybean oil and the like, benzyl benzoate as a solubilizing agent, may be used in combination with benzyl alcohol. Further, a buffering agent, for example, phosphate buffer, sodium acetate buffer, a soothing agent, for example, procaine hydrochloride, a stabilizer such as benzyl alcohol, phenol, may be blended with the antioxidant. The prepared injection is usually, to be filled in an appropriate ampoule.

[0115]

　Administration is oral or parenteral, preferably parenteral administration, in particular, injectable form, nasal dosage form, pulmonary dosage forms, transdermal dosage forms and the like. Examples of injectable dosage forms, for example, intravenous injection, intramuscular injection, intraperitoneal injection, can be administered systemically or locally by subcutaneous injection or the like.

[0116]

　Further, it is possible to select the patient's age, weight, sex, appropriately administration method and symptoms, and the like. The dosage of the antibody or a pharmaceutical composition comprising a polynucleotide encoding an antibody, for example, can be selected in the range of 1000mg from 0.0001mg per body weight 1kg per administration. Alternatively, for example, although the dose can be selected in the range of 0.001 ~ 100000mg / body per patient, not necessarily limited to these numerical values. The dosage, method of administration, patient's weight, age, sex, will vary with conditions like, and can be appropriately selected by those skilled in the art.

[0117]

　Cancer by administering an antibody or said pharmaceutical composition comprising a fragment thereof of the present invention to a subject, preferably a breast cancer, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mast cell tumors, melanoma, adrenocortical cancer, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer or it can be treated and / or preventing head and neck cancer.

[0118]

　Further, a pharmaceutical composition of the present invention, in combination with a pharmaceutical composition comprising an anti-tumor agent or anti-tumor agents as exemplified above, comprising co-administering to a subject, also treatment and / or prevention of cancer It encompassed by the present invention. Antibody or fragment thereof and an antitumor agent of the present invention may be administered simultaneously or separately to the subject. When administered separately, may be even or after be any pharmaceutical composition above, their administration interval, dose, administration route and frequency of administration may be chosen by the specialist. When administered simultaneously, for example, an antibody or fragment thereof and an antitumor agent of the present invention, the pharmaceutical compositions of the pharmaceutical dosage form obtained by mixing formulated in a carrier (or medium) that are pharmacologically acceptable is also included and shall. Also, for any of the above pharmaceutical compositions and dosage forms containing anti-tumor agents, the formulation of the pharmaceutical compositions and dosage forms comprising the antibody of the present invention, formulation, route of administration, dosage, such as cancer description can be applied to.

[0119]

　Accordingly, the present invention comprises administering the pharmaceutical composition of the present invention includes a pharmaceutical composition comprising an anti-tumor agent as exemplified above, cancer treatment and / or a combination medicament for the prevention and it comprising also provides method for the treatment and / or prevention of cancer. Further, the present invention provides an antibody or fragment thereof and an antitumor agent of the present invention, together with pharmacologically acceptable carriers, also provides a pharmaceutical composition for the treatment and / or prevention of cancer.

[0120]

　
　The present invention further provides, DNA encoding the above antibody of the present invention or,, DNA encoding the heavy chain or light chain of said antibody, alternatively, the variable regions of the heavy or light chain of said antibody DNA encoding also provided. Such DNA, for example, in the case of antibody (a), DNA encoding the heavy chain variable region comprising the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 1, 2 and 3, the amino acid sequence of SEQ ID NO: 4, 5 and 6 encoding the light chain variable region comprising the nucleotide sequence encoding the containing DNA, and the like.

[0121]

　Complementarity determining regions encoded by the DNA of these sequences are the regions that determine the specificity of the antibody, except that the antibody regions (i.e., constant regions and framework regions) sequences encoding the other it may be a sequence from an antibody. Here Although the other antibodies, including antibodies from organisms other than human, preferably of human origin in terms of reducing side effects. That is, in the above-described DNA, it is preferred that regions encoding framework regions and each constant region of the heavy and light chains comprise a nucleotide sequence encoding the corresponding amino acid sequences derived from human antibody.

[0122]

　Further, another example of a DNA encoding the antibody of the present invention, for example, DNA encoding the heavy chain variable region comprising the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 8, the region encoding the light chain variable region is SEQ ID NO: DNA comprising the nucleotide sequence encoding the amino acid sequence of 15, and the like. Here, examples of the base sequence encoding the amino acid sequence of SEQ ID NO: 8 is the nucleotide sequence of SEQ ID NO: 23. Further, examples of the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 15 is the nucleotide sequence of SEQ ID NO: 30. Even these DNA, it is preferred that regions encoding the constant region of the heavy and light chains comprise a nucleotide sequence encoding the corresponding amino acid sequences derived from human antibody.

[0123]

　DNA of these antibodies can be obtained, for example, by the method described above or the following method. First, from a hybridoma related to the antibody of the present invention, total RNA was prepared using commercially available RNA extraction kit, and cDNA is synthesized by reverse transcriptase using random primers or the like. Then, in each of the variable regions of known mouse antibody heavy chain gene and light chain gene by PCR method using sequences of the oligonucleotides that are stored respectively in the primers to amplify the cDNA encoding the antibody. The sequence encoding the constant region, a known sequence may be obtained by amplifying by PCR. Nucleotide sequence of DNA, by, for example, incorporated into a plasmid or phage for sequencing, it can be determined by a conventional method.

[0124]

　The present invention further relates to polypeptides and DNA described in the following related to the antibody (i) ~ (xiv) (i) ~ (xv) is also provided.

[0125]

　(I) is selected from the group consisting of the amino acid sequence shown in SEQ ID NO: 1, 2 and 3, the heavy chain CDR polypeptide and DNA encoding the polypeptide.

[0126]

　(Ii) is selected from the amino acid sequence shown in SEQ ID NO: 4, 5 and 6, a light chain CDR polypeptide and DNA encoding the polypeptide.

[0127]

　(Iii) SEQ ID NO: 8 and polypeptide and DNA encoding the polypeptide comprising the amino acid sequence of SEQ ID NO: 15, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 23 and SEQ ID NO: 30.

[0128]

　(Iv) SEQ ID NO: 10 and a DNA encoding the polypeptide and the polypeptide comprising the amino acid sequence of SEQ ID NO: 15, for example SEQ ID NO: 25 and a DNA comprising the nucleotide sequence of SEQ ID NO: 30.

[0129]

　(V) SEQ ID NO: 7 and polypeptide and DNA encoding the polypeptide comprising the amino acid sequence of SEQ ID NO: 15, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 22 and SEQ ID NO: 30.

[0130]

　(Vi) DNA encoding the polypeptide and the polypeptide comprising the amino acid sequence of SEQ ID NO: 8 and SEQ ID NO: 13, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 23 and SEQ ID NO: 28.

[0131]

　(Vii) SEQ ID NO: 7 and polypeptide and DNA encoding the polypeptide comprising the amino acid sequence of SEQ ID NO: 12, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 22 and SEQ ID NO: 27.

[0132]

　(Viii) SEQ ID NO: 8 and polypeptide and DNA encoding the polypeptide comprising the amino acid sequence of SEQ ID NO: 12, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 23 and SEQ ID NO: 27.

[0133]

　(Ix) DNA encoding the polypeptide and the polypeptide comprising the amino acid sequence of SEQ ID NO: 7 and SEQ ID NO: 13, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 22 and SEQ ID NO: 28.

[0134]

　(X) DNA encoding the polypeptide and the polypeptide comprising the amino acid sequence of SEQ ID NO: 10 and SEQ ID NO: 14, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 25 and SEQ ID NO: 29.

[0135]

　(Xi) DNA encoding the polypeptide and the polypeptide comprising the amino acid sequence of SEQ ID NO: 8 and SEQ ID NO: 11, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 23 and SEQ ID NO: 26.

[0136]

　(Xii) SEQ ID NO: 7 and DNA encoding the polypeptide and the polypeptide comprising the amino acid sequence of SEQ ID NO: 11, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 22 and SEQ ID NO: 26.

[0137]

　(Xiii) SEQ ID NO: 9 and DNA encoding the polypeptide and the polypeptide comprising the amino acid sequence of SEQ ID NO: 15, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 24 and SEQ ID NO: 30.

[0138]

　(Xiv) SEQ ID NO: 20 and a DNA encoding the polypeptide and the polypeptide comprising the amino acid sequence of SEQ ID NO: 21, for example, DNA comprising the nucleotide sequence of SEQ ID NO: 31 and SEQ ID NO: 32.

[0139]

　(Xv) wherein (i) ~ (xiv) a polypeptide, or a fragment thereof comprising one or a plurality of amino acid substitutions in the polypeptide to a heavy chain constant region described, as well as the DNA encoding the polypeptide or fragment thereof.

[0140]

　These polypeptides and DNA, as described above, can be prepared using genetic recombination techniques.

[0141]

　
　summarizing the present invention described above to below.

[0142]

　(1) and a light chain variable region comprising the heavy chain complementarity determining regions of the variable region to SEQ ID NO: 4, 5 and 6 including the complementarity determining regions of SEQ ID NO: 1, 2 and 3, and, caprin-1 protein antibody or fragment thereof having the immunological reactivity with.

[0143]

　(2) variable regions of the heavy chain comprises the amino acid sequence of SEQ ID NO: 8, and antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 15 (1).

[0144]

　(3) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 10, and an antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 15 (1).

[0145]

　(4) variable regions of the heavy chain comprises the amino acid sequence of SEQ ID NO: 7 and an antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 15 (1).

[0146]

　(5) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 8, and antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 13 (1).

[0147]

　(6) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 7 and an antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 12 (1).

[0148]

　(7) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 8, and antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 12 (1).

[0149]

　(8) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 7 and an antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 13 (1).

[0150]

　(9) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 10, and an antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 14 (1).

[0151]

　(10) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 8, and antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 11 (1).

[0152]

　(11) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 7 and an antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 11 (1).

[0153]

　(12) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 9 and an antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 15 (1).

[0154]

　(13) the variable region of the heavy chain comprises the amino acid sequence of SEQ ID NO: 20, and an antibody or fragment thereof according to the variable region of the light chain comprising the amino acid sequence of SEQ ID NO: 21 (1).

[0155]

　(14) human antibodies, humanized antibodies, chimeric antibodies, a single chain antibody or a multispecific antibody, antibody or fragment thereof according to (1) to (13).

[0156]

　(15) an antibody or fragment thereof according to the anti-tumor agent conjugated (1) to (13).

[0157]

　(16) comprising said one or more amino acid substitutions in the heavy chain constant region of the antibody, the antibody according to (1) to (15).

[0158]

　(17) wherein the antibody is an antibody obtained by removing fucose bound to N- acetylglucosamine sugar chain reducing end of N- glycoside-linked sugar chain which binds to the heavy chain constant region, described in (1) - (16) antibody of.

[0159]

　(18) a composition of the antibody, fucose is bound to the antibody and the heavy chain constant region to the binding of N- glycoside-linked sugar chain sugar chain reducing end of N- acetyl glucosamine as described in (17) (1 ) - comprising the antibody according to (16), the antibody of the composition.

[0160]

　(19) cells producing a composition of an antibody according to an antibody or (18) described in (17).

[0161]

　(20) (1) an antibody or fragment thereof according to any one of (15), comprise a composition of antibodies as an active ingredient (16) or an antibody according to (17) or (18) and I said, treatment and / or a pharmaceutical composition for the prevention of cancer.

[0162]

　(21) wherein the cancer is breast cancer, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mastocytoma, melanoma, adrenocortical cancer, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer or head and neck cancer, pharmaceutical composition according to (20).

[0163]

　(22) (20) or a pharmaceutical composition according to (21), comprising a pharmaceutical composition comprising an anti-tumor agent, a combination medicament for the treatment and / or prevention of cancer.

[0164]

　(23) (1) ~ (16) DNA encoding an antibody or fragment thereof as described.

[0165]

　Antibody or fragment thereof according to any one of (24) (1) - (16) The antibody according to (17), the composition of the antibody according to (18), according to (20) or (21) the pharmaceutical composition or a combination medicament according to (22), comprising administering to a subject, the treatment and / or prevention of cancer.
Example

[0166]

　Hereinafter, although now be described more concretely by way of the present invention examples, the scope of the present invention shall not be limited these specific examples.

[0167]

　Example 1: Rabbit Preparation of Anti-caprin-1 monoclonal antibodies using
　a mixture of human caprin-1 protein 300μg complete adjuvant equivalent of Freund's prepared in Example 3 of WO2010 / 016526, which rabbit 1 birds per of the antigen solution. The second and subsequent immunization with a mixture with incomplete adjuvant and Freund. After administering the antigen solution intraperitoneally 12-week-old rabbits completed the immunized for 8 doses every 2-3 weeks. Obtain lymphocytes from each spleen was excised from the final immunization after 4 days, with myeloma cells 240E-W2 rabbit 1: was mixed with 2 ratio, there RPMI containing 10% FBS was warmed to 37 ° C. adding medium 200μL and PEG solution prepared by mixing a 800μL of PEG 1500, was performed cell fusion to stand for 5 minutes. After centrifugation to remove the supernatant, the cells were suspended in RPMI medium (HAT selection medium) 300 mL containing 10% FBS plus 2% equivalents of a HAT solution, each 100μL per well of 96-well plates, 80 plates It was seeded. 7 days, 37 ° C., 5 of 5% CO 2 by culturing at conditions, spleen cells with rabbit myeloma cells to obtain a fused hybridomas.

[0168]

　Hybridomas produced were selected hybridomas as an index the reactivity with respect to caprin-1 protein of the antibody produced. 100μL added CAPRIN-1 protein solution 1μg / mL in 96-well plates per well, and allowed to stand for 18 hours at 4 ℃. After 3 times washing each well with PBS-T, allowed to stand at room temperature for 3 hours 0.5% Bovine Serum Albumin (BSA) solution was 400μL added per well. The solution with the exception of, after 3 times washing the wells with 1 per well 400μL of PBS-T, each culture supernatant of the hybridomas obtained above was added 100μL per well and allowed to stand at room temperature for 2 hours. After each well was washed three times with PBS-T, and allowed to stand at room temperature for 1 hour HRP-labeled anti-rabbit antibody diluted 5000 fold with PBS was added per well 100 .mu.L. After washing the wells three times with PBS-T, was chromogenic reaction for 15 to 30 minutes stand to be added 100μL per well of TMB substrate solution. After color development, the reaction was added 100μL per well of 1N sulfuric acid was stopped and the absorbance measured values ​​of 450nm and 595nm using an absorption spectrometer. As a result, a plurality selected hybridomas producing antibodies absorbance value is high.

[0169]

　It was added culture to the plate so as to have a concentration of 0.5 pieces in hybridoma a 96-well plate per well, which were selected. After 1 week, hybridomas forming single colonies in the wells were observed. And further culturing them well of the cell, the cloned hybridomas were selected hybridomas as an index the reactivity with respect to caprin-1 protein of the antibody produced. Result of the reactivity evaluation of caprin-1 protein of each antibody by the same operation as above, to obtain a plurality of hybridoma lines that produce rabbit monoclonal antibody showing reactivity to caprin-1 protein.

[0170]

　Was then selected those that are reactive to human cancer cell surface that express the CAPRIN-1 from rabbit monoclonal antibodies that are reactive to them CAPRIN-1 protein. Specifically, 2 × 10 5 of 5 amino human lung cancer cell line QG56 and human breast cancer cell BT-474 (obtained from ATCC) were centrifuged at microcentrifuge tubes 1.5mL mL, respectively, to which each of the above hybridomas the culture supernatant 100μL was added and allowed to stand on ice for 1 hour. After washing with PBS, PBS containing 0.5% FBS (-) (0.5% FBS -PBS (-)) at 100-fold diluted FITC-labeled anti-rabbit IgG (H + L) antibody or Alexa488-labeled anti-rabbit IgG (h + L) was added, and was allowed to stand on ice for 1 hour. 0.5% FBS-PBS (-) After washing, the cells 0.2μg / mL of Propidium iodide and 0.5% FBS-PBS (-) to the suspended cells, FACSCalibur TM or FACSVerse TM (Becton the fluorescence intensity was measured at Dickinson and Company). On the other hand, the same operation was performed using a medium for hybridoma culture, and a negative control sample. As a result, the fluorescence intensity is stronger than the negative control, i.e., caprin-1 was selected one rabbit anti caprin-1 monoclonal antibodies that react strongly to the cell surface of cancer cells QG56 and BT-474 are expressed.

[0171]

　Next, rabbit anti-caprin-1 monoclonal antibody obtained above, WO2010 / 016526, along with the method described in Example 5 to obtain an amplified fragment of the gene encoding the variable region gene sequences and their amino acid sequence It was analyzed. Specifically mRNA is extracted from hybridomas producing rabbit anti caprin-1 monoclonal antibody, by RT-PCR method using primers specific for the rabbit variable domain sequence, the heavy chain variable (VH) region and the antibody It was to get the gene of the light chain variable (VL) region. And inserts them gene into the cloning vector was determined respective nucleotide sequences according to a conventional method.

[0172]

　From the resulting rabbit anti caprin-1 monoclonal antibody, SEQ ID NO: heavy chain shown in 20 variable regions and CDRl ~ 3 is SEQ ID NO: 1 in the light chain variable region, SEQ ID NO: 2, SEQ ID NO: 3 amino acid sequence becomes, the light chain variable regions and CDRl ~ 3 is SEQ ID NO: 4 in the light chain variable region shown in SEQ ID NO: 21, SEQ ID NO: 5, it was confirmed that the amino acid sequence of SEQ ID NO: 6.

[0173]

　Then the reaction was confirmed of the various to human cancer cells of the acquired rabbit anti-CAPRIN-1 monoclonal antibody. Is a human cancer cells in which the expression of CAPRIN-1 gene have been identified, breast cancer cells (BT-474, MDA-MB -361), colon cancer cells (HT-29), lung cancer cells (QG56), gastric cancer cells ( NCI-N87), uterine cancer cells (HEC-1-A), prostate cancer cells (22Rv1), pancreatic cancer cells (Panc10.5), liver cancer cells (Hep3B), ovarian cancer cells (SKOV3), renal carcinoma cells ( Caki-2), brain tumor cells (U-87MG), bladder cancer cells (T24, HT-1376), esophageal cancer cells (OE33), leukemia cells (OCI-AML5), lymphoma cells (Ramos), gallbladder cancer cells (TGBC14TKB ), fibrosarcoma cells (HT-1080), melanoma cells (G-361), adrenal cortical cancer cells (A-673), Ewing's tumor cells (RD-ES), Hodgkin's lymphoma cells (RPMI1666), mesothelioma cells ( NCI-H2452), multiple myeloma cells (IM-9), testicular cancer cells (NT / D1), in thyroid cancer cells (TT) or head and neck cancer cells (FaDu), reacted to acquired the antibody, flow It was evaluated fluorescence intensity cytometry. In micro centrifuge tubes of 1.5mL capacity Kakugan cells 10, respectively 6 collected in pieces, added the culture supernatant of hybridomas that produce a rabbit anti-CAPRIN-1 monoclonal antibody obtained above the (100μL) to each tube, It was allowed to react for 1 hour at 4 ℃. 0.5% FBS-PBS (-) was washed with, 0.5% FBS-PBS - supplemented with 50-fold diluted FITC-labeled goat anti-rabbit IgG (H + L) antibody (Jackson ImmunoResearch Inc.) () , it was allowed to stand for 60 minutes at 4 ℃. After washing with 0.5% final concentration including the Propidium iodide of 0.2μg / mL FBS-PBS - 0.5 % FBS-PBS () (-) to the suspended cells, FACSCalibur TM or FACSVerse TM ( and the fluorescence intensity was measured using a Becton Dickinson and Company). On the other hand, as a negative control, using a medium for cultivation of a hybridoma to those prepared in the same manner as above to obtain a negative control sample. As a result, in all of the cancer cells used in the above evaluation, the fluorescence intensity in the case of using the culture supernatant of hybridomas producing rabbit anti caprin-1 monoclonal antibody, stronger than the fluorescence intensity in the case of using the negative control It was. These results, rabbit anti-caprin-1 monoclonal antibody, it was confirmed that reacts with caprin-1 cancer cell membrane surface of human cancer cells.

[0174]

　Example 2: Human - rabbit chimeric anti-caprin-1 monoclonal antibodies produced
　in Example 1 for expression of the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 20 was confirmed rabbit anti caprin-1 monoclonal antibody inserting a gene, a gene for expressing a light chain variable region represented by SEQ ID NO: 21, respectively light chain constant region of a mammalian cell expression vector and human IgG1 to the heavy chain constant region was inserted the human IgG1 It was inserted into a mammalian cell expression vector which is. Two recombinant expression vector prepared by introducing into a mammalian cell according to a conventional method person - to obtain a culture supernatant containing - (rabbit chimeric human) rabbit chimeric anti caprin-1 antibody. Hitrap the culture supernatant containing the resulting chimeric antibody according to the conventional method Protein A SepharoseFF (GE Healthcare Co., Ltd.) was purified using, PBS (-) 0.22μm of the filter is replaced in (Millipore) in to prepare what was filtered.

[0175]

　Example 3: Preparation of humanized anti-caprin-1 monoclonal antibodies
　Next, CDRl ~ in CDRl ~ 3 and the light chain variable region in the heavy chain variable region of rabbit anti-caprin-1 monoclonal antibody was confirmed in Example 1 based on the information of the 3 amino acid sequence and nucleotide sequence, heavy chain variable region represented by SEQ ID NO: 7 consisting of heavy chain variable regions of CDRl ~ 3 is SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 amino acid sequence the amino acid sequence was designed nucleotide sequences to allow expression of which was inserted into a mammalian cell expression vector for heavy chain constant region was inserted the human IgG1. Similarly, the base to be able to express the amino acid sequence of the light chain variable region represented by SEQ ID NO: 11 CDRl ~ 3 of the light chain variable region SEQ ID NO: 4, comprising the amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 6 design a sequence, which was inserted into a mammalian cell expression vector for the light chain constant region of human IgG1 has been inserted. The two recombinant expression vector and introduced into mammalian cells according to a conventional method, consisting of the light chain full-length amino acid sequence as shown by the heavy chain full-length amino acid sequence as SEQ ID NO: 11 of SEQ ID NO: 7 humanized antibody # to obtain a culture supernatant containing 0.

[0176]

　Similarly, CDRl ~ 3 is SEQ ID NO: 1 heavy chain variable region, the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 8 comprising the amino acid sequence of SEQ ID NO: 2 and SEQ ID NO: 3, SEQ ID NO: 11 to obtain a culture supernatant containing the humanized antibody # 1 consisting of the amino acid sequence of the light chain variable region represented.

[0177]

　Further in the same manner, to obtain a culture supernatant containing the humanized antibody # 2 to 10 below.

[0178]

　And the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 8, the light which CDRl ~ 3 of the light chain variable region SEQ ID NO: 4, represented by SEQ ID NO: 12 comprising the amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 6 humanized antibody # 2 comprising a strand full-length amino acid sequence.

[0179]

　And the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 8, the light which CDRl ~ 3 of the light chain variable region SEQ ID NO: 4, represented by SEQ ID NO: 13 comprising the amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 6 humanized antibody # 3 comprising a chain full-length amino acid sequence.

[0180]

　SEQ ID NO: amino acid sequence of the heavy chain variable region represented by 7, the humanized antibody comprising a light chain full-length amino acid sequence represented by SEQ ID NO: 12 # 4.

[0181]

　And the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 7, and a light chain full-length amino acid sequence represented by SEQ ID NO: 13 humanized antibody # 5.

[0182]

　And the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 7, a light which CDRl ~ 3 of the light chain variable region SEQ ID NO: 4, represented by SEQ ID NO: 15 comprising the amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 6 humanized antibodies # 6 comprising a chain full-length amino acid sequence.

[0183]

　And the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 8, and a light chain full-length amino acid sequence shown in SEQ ID NO: 15 humanized antibody # 7.

[0184]

　And the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 9 comprising a heavy chain variable region of CDRl ~ 3 is SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 amino acid sequence, light represented by SEQ ID NO: 15 humanized antibody # 8 consisting of a chain full-length amino acid sequence.

[0185]

　Heavy chain variable regions of CDRl ~ 3 is SEQ ID NO: 1, SEQ ID NO: 2 and the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 10 comprising the amino acid sequence of SEQ ID NO: 3, of the light chain variable region CDRl ~ 3 There SEQ ID NO: 4, consisting of a light chain full-length amino acid sequence shown in SEQ ID NO: 14 comprising the amino acid sequence of SEQ ID NO: 5 and SEQ ID NO: 6 humanized antibody # 9.

[0186]

　And the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 10, a humanized antibody # 10 consisting of a light chain full-length amino acid sequence represented by SEQ ID NO: 15.

[0187]

　The resulting culture supernatant containing the humanized antibody # 0 to # 10 was purified using Hitrap Protein A SepharoseFF (GE Healthcare Inc.) according to a conventional method, PBS (-) 0.22μm filter by substituting were prepared that (Millipore).

[0188]

　Example 4: Human - rabbit chimeric antibody, reactivity to antigen specificity and cancer cells of human antibody # 0 to # 10
　were then prepared in Example 2 Human - and rabbit chimeric antibody, prepared in Example 3 the specificity reactivity to CAPRIN-1 protein of humanized antibody # 0 to # 10 according to a conventional method, was confirmed by ELISA. Specifically, 100μL added to a 96-well plate per well of PBS solution containing caprin-1 protein in advance 5 .mu.g / mL, and allowed to stand 18 hours at 4 ° C.. After washing each well with PBS-T, allowed to stand for 3 hours at room temperature blocking solution consisting of PBS solution containing 5% skim milk was added 400μL per well. The solution with the exception of, after washing the wells with PBS-T, the human was adjusted to 1 .mu.g / mL in PBS containing 0.2% skim milk - rabbit chimeric antibodies, the respective solutions containing the humanized antibody # 0 ~ # 10 by 50μL per well was added to each well and allowed to stand at room temperature for 1 hour. As a negative control, were prepared wells was added at the same antibody concentration of human IgG antibody does not react to caprin-1 protein have been identified, and wells without addition of the antibody at the same time. After each well was washed three times with PBS-T, and allowed to stand at room temperature for 1 hour the HRP-labeled anti-human IgG antibody diluted 3000-fold with PBS containing 0.2% skim milk was added 50μL per well . After washing the wells three times with PBS-T, was color reaction to stand for 1 to 30 minutes with TMB substrate solution (Thermo Co.) was added 100μL per well. After color development, the reaction was added 100μL per well of 1N sulfuric acid was stopped and the absorbance measured values ​​of 450nm and 630nm using an absorption spectrometer. At the same time caprin-1 protein wells not a solid phase (non-solid phase wells) be prepared, the measurement was carried out similarly added each antibody. As a result, the absorbance values ​​of the wells supplemented with human IgG antibody does not react to caprin-1 protein used as negative controls have been tested, but were equally low values ​​and the well without addition of the antibody against, the human - absorbance values ​​rabbit chimeric antibodies and humanized antibodies # 0 to # 10 were added respectively wells exhibited equally high value. Further, humans caprin-1 protein relative to wells that are not solid - rabbit chimeric antibodies and humanized antibodies # 0 ~ # 10 showed only negative control comparable absorbance values. This result, human - rabbit chimeric antibodies and humanized antibodies # 0 to # 10 was confirmed to react specifically to caprin-1 protein.

[0189]

　Then, humans react specifically to CAPRIN-1 protein - was confirmed reactivity to rabbit chimeric antibody and humanized antibody # 0 to # 10 of the various types of human cancer cells as well as mouse cancer cells. Is a human cancer cells in which the expression of CAPRIN-1 gene have been identified, breast cancer cells (BT-474, MDA-MB -361), colon cancer cells (HT-29), lung cancer cells (QG56), gastric cancer cells ( NCI-N87), uterine cancer cells (HEC-1-A), prostate cancer cells (22Rv1), pancreatic cancer cells (Panc10.5), liver cancer cells (Hep3B), ovarian cancer cells (SKOV3), renal carcinoma cells ( Caki-2), brain tumor cells (U-87MG), bladder cancer cells (T24, HT-1376), esophageal cancer cells (OE33), leukemia cells (OCI-AML5), lymphoma cells (Ramos), gallbladder cancer cells (TGBC14TKB ), fibrosarcoma cells (HT-1080), melanoma cells (G-361), adrenal cortical cancer cells (A-673), Ewing's tumor cells (RD-ES), Hodgkin's lymphoma cells (RPMI1666), mesothelioma cells ( NCI-H2452), multiple myeloma cells (IM-9), testicular cancer cells (NT / D1), in thyroid cancer cells (TT) or head and neck cancer cells (FaDu), purified human - rabbit chimeric antibody and humanized antibodies # 0 to # 10 was reacted respectively, were evaluated fluorescence intensity by flow cytometry. Specifically, the micro-centrifuge tube of 1.5mL capacity Kakugan cells 5 × 10 each 5 gathered one by one, person - a final concentration of rabbit chimeric antibody and humanized antibody # 0 to # 10 is 50μg / mL as described above, it was added to each tube and allowed to react for 1 hour at 4 ℃. 0.5% FBS-PBS (-) was washed twice with, 0.5% FBS-PBS - an Alexa488-labeled goat anti-human IgG diluted 100 times with (H + L) (manufactured by Life Technologies, Inc.) antibodies () It was allowed to stand for 60 minutes in addition to 4 ℃. After washing with 0.5% final concentration including the Propidium iodide of 0.2μg / mL FBS-PBS - 0.5 % FBS-PBS () (-) to the suspended cells, FACSCalibur TM or FACSVerse TM ( and the fluorescence intensity was measured using a Becton Dickinson and Company). On the other hand, as a negative control, using a medium for hybridoma culture was used to prepare in the same manner as above. As a result, in all of the cancer cells used in the above evaluation, human - fluorescence intensity of rabbit chimeric antibodies and humanized antibodies # 0 to # 10 was stronger than the fluorescence intensity in the case of using the negative control. These results, human - rabbit chimeric antibodies and humanized antibodies # 0 ~ # 10, it was confirmed that reacts with caprin-1 protein expressed on human cancer cell membrane surface.

[0190]

　Example 5: Human - Antitumor activity against various human cancer cell rabbit chimeric antibodies and humanized antibodies # 0 ~ # 10
　Next, human prepared in Example 2 - and rabbit chimeric antibodies, human prepared in Example 3 the anti-tumor effects on various types of human cancer cells in the antibody # 0 to # 10 was evaluated by the ADCC activity.

[0191]

　Human - as rabbit chimeric antibody and comparing antibody against humanized antibody # 0 ~ # 10, used the following anti-caprin-1 antibody.

[0192]

　WO2010 / 016526 Comparative Antibody 1 having a light chain variable region of the heavy chain variable region SEQ ID NO: 27 SEQ ID NO: 26 in the document be an antibody described, the heavy chain variable region SEQ ID NO: 29 SEQ ID NO: 28 comparison antibodies 2 having a light chain variable region, the light chain variable region of comparison antibody 3, the heavy chain variable region of SEQ ID NO: 32 and SEQ ID NO: 33 having a light chain variable region of the heavy chain variable region SEQ ID NO: 31 SEQ ID NO: 30 Comparative antibodies with 4, comparative antibody having a heavy chain variable region and comparing antibody 5 having a light chain variable region of SEQ ID NO: 35, the light chain variable region of the heavy chain variable region SEQ ID NO: 37 SEQ ID NO: 36 SEQ ID NO: 34 6, the heavy chain variable comparison antibodies 7 having a light chain variable region of a region between SEQ ID NO: 39, comparing antibody 8 having a light chain variable region of the heavy chain variable region SEQ ID NO: 41 SEQ ID NO: 40 SEQ ID NO: 38, SEQ ID NO: comparison antibodies 9 having a heavy chain variable regions of the variable region and SEQ ID NO: 43 of 42, heavy chain comparison antibody 10, SEQ ID NO: 46 having a light chain variable region of the heavy chain variable region SEQ ID NO: 45 SEQ ID NO: 44 comparison antibody 11 having light chain variable regions of the variable region to SEQ ID NO: 47.

[0193]

　WO2011 / 096517 an antibody as described in comparative antibody 12 having light chain variable region of the heavy chain variable region SEQ ID NO: 47 SEQ ID NO: 43 in the same literature, the heavy chain variable region SEQ ID NO: 53 SEQ ID NO: 43 comparison antibody 13 having light chain variable region.

[0194]

　WO2011 / 096528 an antibody as described in comparative antibody 14 having light chain variable region of the heavy chain variable region SEQ ID NO: 47 SEQ ID NO: 43 in the same literature, the heavy chain variable region SEQ ID NO: 55 SEQ ID NO: 51 comparison antibody 15 having light chain variable region, a heavy chain comparison antibody 16 having light chain variable regions of the variable region to SEQ ID NO: 63, the light chain variable region of the heavy chain variable region SEQ ID NO: 80 SEQ ID NO: 76 SEQ ID NO: 59 comparison antibody 17 having, compared antibody having a heavy chain variable region and compared with a light chain variable region of SEQ ID NO: 88 antibody 18 light chain variable region of the heavy chain variable region SEQ ID NO: 96 SEQ ID NO: 92 SEQ ID NO: 84 19.

[0195]

　WO2011 / 096519 an antibody as described in comparative antibody 20 having light chain variable region of the heavy chain variable region SEQ ID NO: 46 SEQ ID NO: 42 in the document.

[0196]

　WO2011 / 096533 an antibody as described in comparative antibody 21 having light chain variable region of the heavy chain variable region SEQ ID NO: 51 SEQ ID NO: 43 in the same literature, the heavy chain variable region SEQ ID NO: 51 SEQ ID NO: 47 comparison antibody 23 having light chain variable region of comparison antibody 22, a heavy chain variable region SEQ ID NO: 67 SEQ ID NO: 63 having a light chain variable region.

[0197]

　WO2011 / 096534 comparative antibody 24 having light chain variable region of the heavy chain variable region SEQ ID NO: 47 SEQ ID NO: 43 in the document be an antibody described, the heavy chain variable region SEQ ID NO: 51 SEQ ID NO: 43 comparison antibody 26 having light chain variable region of comparison antibody 25, a heavy chain variable region of SEQ ID NO: 63 and SEQ ID NO: 67 having a light chain variable region.

[0198]

　WO2013 / 018894 comparative antibody 27 having light chain variable region of the heavy chain variable region SEQ ID NO: 13 SEQ ID NO: 9 in the literature an antibody as described in, the heavy chain variable region SEQ ID NO: 23 SEQ ID NO: 19 comparison antibody 28 having light chain variable region, the light chain variable region of the heavy chain comparison antibody 29 having light chain variable regions of the variable region to SEQ ID NO: 53, a heavy chain variable region SEQ ID NO: 62 SEQ ID NO: 58 SEQ ID NO: 9 comparison antibody 30 having, comparison antibodies having the heavy comparison antibody 31 having light chain variable region of chain variable region and SEQ ID NO: 65, the light chain variable region of the heavy chain variable region SEQ ID NO: 73 SEQ ID NO: 69 SEQ ID NO: 63 32, compared with a light chain variable region of the heavy chain variable region SEQ ID NO: 81 SEQ ID NO: 77 antibody 33.

[0199]

　WO2013 / 018892 an antibody as described in comparative antibody 34 having light chain variable region of the heavy chain variable region SEQ ID NO: 12 SEQ ID NO: 8 in the literature.

[0200]

　WO2013 / 018891 an antibody as described in comparative antibody 35 having light chain variable region of the heavy chain variable region SEQ ID NO: 12 SEQ ID NO: 8 in the literature.

[0201]

　WO2013 / 018889 an antibody as described in comparative antibody 36 having light chain variable region of the heavy chain variable region SEQ ID NO: 12 SEQ ID NO: 8 in the literature.

[0202]

　WO2010 / 018883 an antibody as described in comparative antibody 37 having light chain variable region of the heavy chain variable region SEQ ID NO: 12 SEQ ID NO: 8 in the literature.

[0203]

　WO2013 / 125636 an antibody as described in comparative antibody 38 having the heavy chain variable regions of the variable region and SEQ ID NO: 7 SEQ ID NO: 6 in the literature.

[0204]

　WO2013 / 125654 an antibody as described in comparative antibody 39 having light chain variable region of the heavy chain variable region SEQ ID NO: 54 SEQ ID NO: 52 in the document. Comparison antibody 40 having light chain variable region of the heavy chain variable region SEQ ID NO: 23 SEQ ID NO: 21 in the document. Comparison antibody 41 having light chain variable region of the heavy chain variable region SEQ ID NO: 23 SEQ ID NO: 25 in the document. Comparison antibody 42 having light chain variable region of the heavy chain variable region SEQ ID NO: 18 SEQ ID NO: 16 in the document. Comparison antibody 43 having light chain variable region of the heavy chain variable region SEQ ID NO: 33 SEQ ID NO: 29 in the document. Comparison antibody 44 having light chain variable region of the heavy chain variable region SEQ ID NO: 43 SEQ ID NO: 39 in the document. Comparison antibody 45 having light chain variable region of the heavy chain variable region SEQ ID NO: 43 SEQ ID NO: 49 in the document.

[0205]

　WO2013 / 125630 an antibody as described in comparative antibody 46 having light chain variable region of the heavy chain variable region SEQ ID NO: 15 SEQ ID NO: 11 in the document.

[0206]

　WO2013 / 125640 an antibody as described in comparative antibody 47 having light chain variable region of the heavy chain variable region SEQ ID NO: 15 SEQ ID NO: 11 in the document. Comparison antibody 48 having light chain variable region of the heavy chain variable region SEQ ID NO: 25 SEQ ID NO: 21 in the document.

[0207]

　Incidentally, the antibody (Comparative antibody 1-48) were compared, the heavy chain variable and gene for expressing the amino acid sequences of the region, and a gene for expressing a light chain variable region, a heavy chain constant for each human IgG1 region inserted mammalian cell expression vector pcDNA4 / myc-His (Life Technologies Inc.) and mammalian cell expression vector for light chain constant region was inserted the human IgG1 pcDNA3.1 / myc-His (Life Technologies, Inc. was inserted into), purified two recombinant expression vector prepared using Hitrap human chimeric reduction or humanized antibodies obtained by introducing into mammalian cells Protein a SepharoseFF (GE Healthcare Inc.) according to a conventional method, PBS (-) was used and filtered through a 0.22μm filter (Millipore) by substituting.

[0208]

　Further, as a negative control, wells were added isotype control antibody reacts antibody to wells and caprin-1 protein without the addition of the antibodies not reactive with human carcinoma cell surface caprin-1 is expressed It was prepared adding wells. Each antibody final concentration were added to V-bottom 96-well plate so that 5μg / mL.

[0209]

　Effector cells, using human NK cells isolated using conventional methods from human peripheral blood mononuclear cells. Using peripheral blood gravity separation liquid Histopaque of mononuclear cell separation (Sigma-Aldrich) separating the human peripheral blood mononuclear cells, FITC fluorochrome labeled antibody (anti-human CD3 antibody, anti-human CD20 antibody, anti-human CD19 antibody, anti-human CD11c antibody, using reacted with anti-HLA-DR antibody (Pharmingen)) in cell sorter (FACS Vantage SE (Becton, Dickinson and Company)), does not stain with the antibody, NK those separated cell population containing cells, or used was separated using human NK cell isolation kit (Miltenyi Inc.). Human NK cells in V-bottom 96-well plates supplemented with the respective antibodies, 0.4 ~ 2.0 × 10 per well 5 were prepared have been added pieces.

[0210]

　Ha target cells, breast cancer cells (BT-474, MDA-MB -361), colon carcinoma (HT-29), lung cancer cells (QG56), gastric cancer cells (NCI-N87), uterine cancer (HEC-1-A ), before the legislature adenocarcinoma cells (22Rv1), pancreas Zang cancer (Panc10.5), liver Zang cancer cells (Hep3B), egg Chao cancer cells (SKOV3), renal cell carcinoma (Caki-2), Nao neoplasm cells (U-87MG), bladder cancer cells (T24, HT-1376), esophageal cancer cells (OE33), leukemia cells (OCI-AML5), ri nn pa swollen cells (Ramos), bile Nang cancer (TGBC14TKB), line-dimensional flesh swollen cells (HT-1080),メGetting Techno have ma cells ( G-361), adrenal cortical carcinoma (A-673), yuーイnn Corning neoplasm (RD-ES),ホji ki nn ri nn pa swollen (RPMI1666), in mesothelioma (NCI-H2452), multi Requested Procedure bone marrow edema (IM-9), testicular cancer ( NT / D1), thyroid cancer (TT) and wa head and neck cancer (FaDu) with i ta wo. 10 . 6 lastのremember Hikaru Suites cancer cell lines woそme cryぞme cry 50mL capacityのtelecentric chiューbu ni set Circular, 100μCiのku ro ミウRousseau 51 (AiluropodaーDaikin e ru ma have Inc.) wo plusえ37 ℃で1 Time DIC CorporationュベーSuites shi ta. その10% postのFBS wo containingむRPMI1640 culture toで3 back wash shi, before the mindでEito fuェku center have cell na raびni each antibody wo add shi ta 96 well V bottom pu RitzーSuites ni, 1 hole thou ta ri 2 × 10 3 Ge zu zu add shi te, 37 ℃, 5% CO. 2での4 time under conditions of anti Applied sa se ​​ta. After the reaction Applied, disability wo by ke ta cancer kara release Connecticut made ​​me cry ru supernatantのku ro ミウRousseau 51 supernatant wo containing san da supernatant wo eachウェHikaru 50μL zu zu recovery shi te,ウェHikaru bottom surface ni individualシnn chi RitzーTatari ga Uーte I nn Corning Connecticut re ta, LumaPlate- 96 (AiluropodaーDaikin e ru ma have manufactured) ni add shi, dried sa se ​​te, disability wo by ke ta cancer kara release Connecticut made ​​me cry ru supernatantのku ro ミウRousseau 51のamount wo measured shi, anti CAPRIN-1 antibody ni yo ru cancer ni Dui su ru anti-neoplasm effect wo shi ta calculated.

[0211]

　As a result, breast cancer cells (BT-474) Humanized antibodies # 7 for humanized antibody # 10, humanized antibodies # 6 antibodies show the antitumor effect of more than 54%, the humanized antibody # 3, human antibody # 4, the humanized antibody # 2, humanized antibody # 5 represents the activity of 50% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, human against the antibody # 8 showed activity greater than 46%, compared antibody 1-48 is in all 25% or less, the negative control group was 10% or less none.

[0212]

　Breast cancer cells (MDA-MB-361) Humanized antibodies # 7 for humanized antibody # 10, humanized antibodies # 6 antibodies show the antitumor effect of more than 52%, the humanized antibody # 3, humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 showed 45% more active, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized whereas antibody # 8 showed activity greater than 40%, compared antibody 1-48 is in all 25% or less, negative control group was either 6% or less.

[0213]

　Colon cancer cells (HT-29) The humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of more than 43%, the humanized antibody # 3, a humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 represents the activity of 40% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed activity greater than 35%, compared antibody 1-48 are all equal to or lower than 20%, the negative control group was 3% none.

[0214]

　Humanized antibody # 7 for lung cancer cells (QG56), humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of more than 46%, the humanized antibody # 3, the humanized antibody # 4, human antibody # 2, humanized antibody # 5 represents the activity of more than 42%, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 38 whereas showed% more active, compared antibody 1-48 are all equal to or lower than 22%, the negative control group were both 10% or less.

[0215]

　Gastric cancer cell humanized antibody # 7 for (NCI-N87), a humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 45% or more, the humanized antibody # 3, humanized antibodies # 4 , humanized antibody # 2, shows a humanized antibody # 5 over 38% activity, antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 34 whereas showed% more active, compared antibody 1-48 is below all of 15%, the negative control group was 8% or less none.

[0216]

　Uterine cancer cells (HEC-1-A) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of more than 52%, the humanized antibody # 3, humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 showed 45% more active, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized whereas antibody # 8 showed activity greater than 40%, compared antibody 1-48 are all equal to or lower than 20%, negative control group was 5% or less none.

[0217]

　Prostate cancer cells (22Rv1) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of more than 49%, the humanized antibody # 3, Antibody # 4, humanized antibody # 2, humanized antibody # 5 showed 45% more active, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 38% whereas it showed more active, compared antibody 1-48 are all equal to or lower than 20%, negative control group was 12% or less none.

[0218]

　Pancreatic cancer cells (Panc10.5) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 35% or more, the humanized antibody # 3, a humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 represents the activity of 30% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed activity greater than 24%, compared antibody 1-48 is below all 10%, any negative control group was 2% or less.

[0219]

　Hepatoma cells (Hep3B) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 28% or more, the humanized antibody # 3, the humanized antibody # 4, humanized antibody # 2, humanized antibody # 5 represents the activity of 25% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas it showed 21% more active, compared antibody 1-48 are all equal to or lower than 12%, negative control group was 5% or less none.

[0220]

　Ovarian cancer cells (SKOV3) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 35% or more, the humanized antibody # 3, the humanized antibody # 4, humanized antibody # 2, humanized antibody # 5 represents the activity of more than 31%, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas it showed 27% more active, compared antibody 1-48 is below all of 15%, negative control group was 5% or less none.

[0221]

　Renal carcinoma cells (Caki-2) a humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 37% or more, the humanized antibody # 3, a humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 showed 33% more active, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8, whereas showed a 26% more active, compared antibody 1-48 is below all of 15%, negative control group was 5% or less none.

[0222]

　Brain tumor cells (U-87MG) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of more than 36%, the humanized antibody # 3, humanized antibodies # 4 , humanized antibody # 2, humanized antibody # 5 represents the activity of more than 29%, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed activity greater than 24%, compared antibody 1-48 is below all 10%, negative control group was either 6% or less.

[0223]

　Bladder cancer cells (T24) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of more than 36%, the humanized antibody # 3, the humanized antibody # 4, humanized antibody # 2, humanized antibody # 5 showed 33% more active, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas it showed more than 30% of the activity, compared antibody 1-48 is below all of 15%, negative control group was either 6% or less.

[0224]

　Bladder cancer cells (HT-1376) Humanized antibodies # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 45% or more, the humanized antibody # 3, a humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 represents the activity of 40% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed activity greater than 28%, compared antibody 1-48 are all equal to or lower than 20%, negative control group was either 7% or less.

[0225]

　Esophageal cancer cells (OE33) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 35% or more, the humanized antibody # 3, the humanized antibody # 4, humanized antibody # 2, humanized antibody # 5 showed 33% more active, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas it showed more than 30% of the activity, compared antibody 1-48 is below all of 15%, negative control group was either 6% or less.

[0226]

　Leukemia cells (OCI-AML5) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 20% or more, the humanized antibody # 3, humanized antibodies # 4 , humanized antibody # 2, shows a humanized antibody # 5 18% more active, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed activity greater than 15%, compared antibody 1-48 is below all 10%, negative control group was either 6% or less.

[0227]

　Lymphoma cells (Ramos) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 20% or more, the humanized antibody # 3, the humanized antibody # 4, human antibody # 2, humanized antibody # 5 showed 18% more active, antibody # 9, antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 of more than 15% active whereas showed, compared antibody 1-48 is below all 10%, negative control group was either 6% or less.

[0228]

　Humanized antibody # 7 for gallbladder cancer cells (TGBC14TKB), humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 35% or more, the humanized antibody # 3, the humanized antibody # 4, human antibody # 2, humanized antibody # 5 represents the activity of 30% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 25 whereas showed% more active, compared antibody 1-48 is below all of 15%, negative control group was either 6% or less.

[0229]

　Humanized antibody # 7 against fibrosarcoma cells (HT-1080), a humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 30% or more, the humanized antibody # 3, Antibody # 4, humanized antibody # 2, humanized antibody # 5 represents the activity of 25% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas it showed more than 20% of the activity, compared antibody 1-48 is below all 10%, negative control group was either 6% or less.

[0230]

　Melanoma (G-361) Humanized antibodies # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 25% or more, the humanized antibody # 3, the humanized antibody # 4, humanized antibody # 2, humanized antibody # 5 represents the activity of more than 21%, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas it showed 15% more active, compared antibody 1-48 are all equal to or lower than 8%, negative control group was either 6% or less.

[0231]

　Adrenocortical carcinoma cells (A-673) Humanized antibodies # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 50% or more, the humanized antibody # 3, a humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 represents the activity of more than 46%, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed activity greater than 40%, compared antibody 1-48 are all equal to or lower than 20%, the negative control group was 8% or less none.

[0232]

　Ewing's tumor cells (RD-ES) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 48% or higher, the humanized antibody # 3, a humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 represents the activity of 40% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed activity greater than 31%, compared antibody 1-48 is below all of 15%, negative control group was either 6% or less.

[0233]

　Hodgkin lymphoma cell (RPMI1666) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 40% or more, the humanized antibody # 3, the humanized antibody # 4, humanized antibody # 2, humanized antibody # 5 represents the activity of more than 36%, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed more than 30% of the activity, compared antibody 1-48 are all equal to or lower than 20%, negative control group was 5% or less none.

[0234]

　Mesothelioma cells humanized antibody # 7 for (NCI-H2452), humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 35% or more, the humanized antibody # 3, a humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 represents the activity of more than 39%, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed activity greater than 31%, compared antibody 1-48 is below all 10%, negative control group was 5% or less none.

[0235]

　Multiple myeloma cells (IM-9) The humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 35% or more, the humanized antibody # 3, humanized antibody # 4, antibody # 2, humanized antibody # 5 represents the activity of 30% or more, the humanized antibody # 9, antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas it showed 27% more active, compared antibody 1-48 is below all 10%, negative control group was either 6% or less.

[0236]

　Testicular cancer cells (NT / D1) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 37% or more, the humanized antibody # 3, a humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 represents the activity of 30% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas showed activity greater than 25%, compared antibody 1-48 are all equal to or lower than 11%, negative control group was 5% or less none.

[0237]

　Thyroid carcinoma cells (TT) humanized antibody # 7 for humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of more than 42%, the humanized antibody # 3, the humanized antibody # 4, humanized antibody # 2, humanized antibody # 5 represents the activity of 35% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, humanized antibodies # 8 whereas it showed more than 30% of the activity, compared antibody 1-48 is below all of 15%, negative control group was 5% or less none.

[0238]

　For head and neck cancer cells (FaDu), humanized antibody # 7, the humanized antibody # 10, humanized antibody # 6 represents the antitumor effect of 50% or more, the humanized antibody # 3, a humanized antibody # 4, the humanized antibody # 2, humanized antibody # 5 represents the activity of 40% or more, the humanized antibody # 9, a humanized antibody # 1, the humanized antibody # 0, a human - rabbit chimeric antibodies, antibody # 8 whereas it showed more than 35% of the activity, compared antibody 1-48 are all equal to or lower than 20%, the negative control group was 8% or less none.

[0239]

　From the above results, the humanized antibody # 0 ~ # 10, and human - rabbit chimeric antibodies, breast cancer, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, melanoma, adrenocortical cancer, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, to thyroid cancer or head and neck cancer, compared to to exhibit significantly stronger anti-tumor effect than antibodies revealed.

[0240]

　In addition, a humanized antibody # 0 to # 10, and human - rabbit chimeric antibodies, WO2010 / 016526, WO2011 / 096517, WO2011 / 096528, WO2011 / 096519, WO2011 / 096533, WO2011 / 096534, WO2011 / 096535, WO2013 / 018886, WO2013 / 018894, WO2013 / 018892, WO2013 / 018891, WO2013 / 018889, WO2013 / 018883, WO2013 / 125636, WO2013 / 125654, WO2013 / 125630, WO2013 / 125640, WO2013 / 147169, all as described in example of WO2013 / 147176 than antibodies against caprin-1, breast cancer described above, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer , gall bladder cancer, sarcoma, melanoma, adrenocortical cancer, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, showed a significantly stronger anti-tumor activity against multiple myeloma, testicular cancer, thyroid cancer or head and neck cancer.

[0241]

　Incidentally, the anti-tumor effect, an antibody against caprin-1, by mixing effector cells and target cells that have incorporated the chromium 51 was cultured for 4 hours, and measuring the amount of chromium 51 released into the medium after culture, the following formula * is the result showing cytotoxic activity against cancer cell lines was calculated by.

[0242]

　* Formula: cytotoxic activity (%) = - were added (chromium 51 release amount from the target cell at the time of addition of antibody and effector cells to CAPRIN-1 chromium 51 spontaneous release amount from the target cell) ÷ (1N hydrochloric acid chromium 51 release amount from the target cell - chromium 51 spontaneous release amount from the target cell) × 100.

[0243]

　Example 6-1: Production of amino acid substitutions human anti caprin-1 monoclonal antibody in the heavy chain constant region
　humanized antibody # 0 obtained in Example 3, # 2, # 3, # 4, # 5 , # 6, # 7, # 8, # 9, the anti-caprin-1 antibody (hereinafter breaks I having a heavy chain constant region as set forth in SEQ ID NO: 33 which partially acids are substituted in the heavy chain constant region of # 10 We fabricated to as a type anti-CAPRIN-1 antibody). Synthesizing DNA encoding the heavy chain amino acid sequence having a heavy chain variable region represented by SEQ ID NO: 7 and said heavy chain constant region which was inserted into a mammalian cell expression vector according to conventional methods. Moreover, DNA encoding the amino acids of the light chain variable region represented by SEQ ID NO: 11 were prepared which genes encoding the light chain constant region of human IgG1 has been inserted into the insertion mammalian cell expression vector. Two recombinant expression vector prepared was introduced into mammalian cells according to a conventional method to obtain a culture supernatant of the modified type I anti-caprin-1 antibody # 0 humanized antibody # 0. Further Example 3 Humanized Antibody # 1 described in, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, modified I-type in the same manner as described above applies to # 10 anti-caprin-1 antibody # 1, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, to obtain respectively a culture supernatant containing the # 10. Each culture supernatant containing the resulting modified type I anti-caprin-1 antibody # 0 to # 10 was purified using Hitrap Protein A SepharoseFF (GE Healthcare Inc.) according to a conventional method, PBS - replaced with () It was prepared which was filtered through a 0.22μm filter (Millipore) on.

[0244]

　Then Example 3 Humanized antibody # 0 were obtained, the # 1, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, in the heavy chain constant region of # 10 some amino acids was produced in the anti-caprin-1 antibody having a heavy chain constant region of SEQ ID NO: 34 wherein substituted (hereinafter referred to as modified type II anti-caprin-1 antibody). Synthesizing DNA encoding the heavy chain amino acid sequence having a heavy chain variable region represented by SEQ ID NO: 7 and said heavy chain constant region which was inserted into a mammalian cell expression vector according to conventional methods. Also, prepared above, those DNA encoding the amino acids of the light chain variable region represented by SEQ ID NO: 11 was inserted into a mammalian cell expression vector for the gene encoding the light chain constant region was inserted the human IgG1 Got ready. These two recombinant expression vector and introduced into mammalian cells according to a conventional method to obtain a culture supernatant of the modified type II anti-caprin-1 antibody # 0. Further Example 3 Humanized Antibody # 1 described in, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, # modified type II in the same manner as described above for 10 anti-caprin-1 antibody # 1, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, to obtain respectively a culture supernatant containing the # 10. Each culture supernatant containing the resulting modified type II anti-caprin-1 antibody # 0 to # 10 was purified using Hitrap Protein A SepharoseFF (GE Healthcare Inc.) according to a conventional method, PBS - replaced with () It was prepared which was filtered through a 0.22μm filter (Millipore) on.

[0245]

　Example 6-2: Out of the total N- glycoside-linked sugar chain that binds to the heavy chain constant region, the production of anti-CAPRIN-1 antibody having a sugar chain in which fucose to N- acetylglucosamine in the reducing end in the sugar chains is not bound
　then, the humanized antibody # 0 obtained in example 3, # 1, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, the heavy chain constant region of # 10 out of the total N- glycoside-linked sugar chains that bind to, be described as anti-CAPRIN-1 antibody (or less modified type III anti-CAPRIN-1 antibody having a sugar chain in which fucose to N- acetylglucosamine in the reducing end in the sugar chains is not bound ) was obtained by the following method. Is an enzyme that does not catalyze the reaction to convert GDP-6- deoxy -D- Rikiso-4-hexulose to the GDP-L- fucose according to the conventional method GDP-6- deoxy -D- Rikiso-4-hexane slow-reductase (RMD ) mammalian cell expression vector containing a neomycin resistance gene incorporating the gene transfer reagent FreeStyle gene TM　were introduced into mammalian cell lines CHO cells using MAX reagent (Life techonologies). Was prepared STABLE pool of CHO cells RMD is expressing CHO cells transfected with said gene was cultivated in inclusive culture solution G-418. RMD has seven cloned CHO cells constitutively expressed by ultra-dilution method from this stable pool. The RMD amount of gene expression in cloned seven each RMD-CHO cells were evaluated three times a week every other quantitative PCR method, CHO cells RMD gene is permanently stable expression of the (RMD-CHO cells) selection was. Permanently into the RMD-CHO cells expressing RMD, in the same manner as in Example 3, the gene encoding the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 7, a light chain of SEQ ID NO: 11 and a gene encoding the amino acid of the variable region, according to their respective mammalian cell expression vector for light chain constant region was inserted in the mammalian cell expression vector and human IgG1 to the heavy chain constant region was inserted conventional methods of human IgG1 It was introduced to obtain a culture supernatant containing modified type III anti-caprin-1 antibody # 0. Further Example 3 Humanized Antibody # 1 described in, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, # modified type III in the same manner as described above for 10 anti-caprin-1 antibody # 1, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, to obtain respectively a culture supernatant containing the # 10. Each culture supernatant containing the resulting modified type III anti-caprin-1 antibody # 0 to # 10 was purified using Hitrap Protein A SepharoseFF (GE Healthcare Inc.) according to a conventional method, PBS - replaced with () to obtain an antibody composition comprising a modified type III anti-caprin-1 antibody # 0 to prepare those filtered with 0.22μm filter (Millipore) on. It was obtained antibody purification comprising a modified type III anti-caprin-1 antibody # 1 to # 10 in a similar manner. Out of the total N- glycoside-linked sugar chain that binds to the purified heavy chain constant region to be included in these antibody compositions were, anti-CAPRIN having a sugar chain in which fucose to N- acetylglucosamine in the reducing end in the sugar chains is not bound the ratio of 1 antibody was LabChip (registered trademark) GXII (PerkinElmer, Inc.) evaluation result both 80% or more.

[0246]

　Above the RMD-CHO cells, in the same manner as in Example 3, the gene encoding the amino acid sequence of the heavy chain variable region represented by SEQ ID NO: 7, encoding the amino acids of the light chain variable region represented by SEQ ID NO: 11 a gene, a mammalian cell expression vector containing each of the hygromycin resistance gene heavy chain constant region is a light chain constant region of a mammalian cell expression vector and human IgG1 containing hygromycin resistance gene inserted was inserted the human IgG1 the said cells introduced by a conventional method, respectively, and cultured in containing hygromycin B broth was prepared STABLE pool expressing modified type III anti-caprin-1 antibody # 0. Break III anti-CAPRIN-1 antibody # 0 to produce a permanently stable expression to cells in limiting dilution method from this stable pool. Out of the total N- glycoside-linked sugar chain in which each cell is linked to a heavy chain constant region that is included in the antibody composition that has been purified, including the modified type III anti-CAPRIN-1 antibody # 0 to # 10 to produce, sugar reduction the proportion of anti-caprin-1 antibodies at the ends of the N- acetylglucosamine having a sugar chain in which fucose is not bound LabChip (registered trademark) GXII was (PerkinElmer, Inc.) evaluation result both 80% or more.

[0247]

　Example 6-3: the substituted amino acid in the heavy chain constant region, and among all the N- glycoside-linked sugar chain that binds to the heavy chain constant region, bound fucose to N- acetylglucosamine in the reducing end in the sugar chains Preparation of anti-caprin-1 antibody with no carbohydrate
　Next, a humanized antibody # 0 described in example 3, # 1, # 2, # 3, # 4, # 5, # 6, # 7, # 8 , # 9, a heavy chain portion of the constant region amino acid has a heavy chain constant region sequences ID 34 wherein replacement, and total N- glycoside-linked sugar chain which binds to the heavy chain constant region of an antibody of # 10 of, it was produced in the anti-CAPRIN-1 antibody having a sugar chain in which fucose to N- acetylglucosamine in the reducing end in the sugar chains is not bound (hereinafter referred to as modified IV anti-CAPRIN-1 antibody). EXAMPLE to RMD-CHO cells expressing the prepared constitutively GDP-6- deoxy -D- Rikiso 4- hexane slow-reductase (RMD) at 6-2, mutant heavy Kusarijo prepared in Example 6-1 a mammalian cell expression vector for synthesizing this DNA encoding the heavy chain amino acid sequences were inserted in a conventional manner having a heavy chain variable region of human IgG1 represented by SEQ ID NO: 7 and a normal region, represented by SEQ ID NO: 11 that the light chain variable region amino acid to synthesize DNA encoding the breaks type IV by introducing mammalian cell expression vector for the gene encoding the amino acids of the light chain constant region was inserted the human IgG1 humanized antibody # 0 to give the anti-CAPRIN-1 culture supernatant of antibody # 0. Further Example 3 Humanized Antibody # 1 described in, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, reforming type IV in the same manner as described above applies to # 10 anti-caprin-1 antibody # 1, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, to obtain respectively a culture supernatant containing the # 10. Each culture supernatant containing the resulting modified IV anti-caprin-1 antibody # 0 to # 10 was purified using Hitrap Protein A SepharoseFF (GE Healthcare Inc.) according to a conventional method, PBS - replaced with () to obtain an antibody composition comprising a modified IV anti-caprin-1 antibody # 0 and filtered through a 0.22μm filter (Millipore) on. It was obtained antibody purification comprising a modified IV anti-caprin-1 antibody # 1 to # 10 in a similar manner. Out of the total N- glycoside-linked sugar chain that binds to the purified heavy chain constant region to be included in these antibody compositions were, anti-CAPRIN having a sugar chain in which fucose to N- acetylglucosamine in the reducing end in the sugar chains is not bound the ratio of 1 antibody was LabChip (registered trademark) GXII (PerkinElmer, Inc.) evaluation result both 80% or more.

[0248]

　Hygromycin which this synthesizing DNA encoding the heavy chain amino acid sequence having a heavy chain variable region of human IgG1 represented by SEQ ID NO: 7 and mutant heavy chain constant region produced in Example 6-1 was inserted in a conventional manner a mammalian cell expression vector containing the puromycin resistance gene, was synthesized DNA encoding amino acids of the light chain variable region represented by SEQ ID NO: 11, a gene encoding the amino acids of the light chain constant region of human IgG1 was inserted the cells transfected with mammalian cell expression vector containing the hygromycin resistance gene, and cultured in containing hygromycin B broth was prepared STABLE pool expressing modified IV anti-caprin-1 antibody # 0. Breaks IV anti-CAPRIN-1 antibody # 0 to produce a permanently stable expression to cells in limiting dilution method from this stable pool. Further Example 3 Humanized Antibody # 1 described in, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, reforming type IV in the same manner as described above applies to # 10 anti-caprin-1 antibody # 1, # 2, # 3, # 4, # 5, # 6, # 7, # 8, # 9, was produced constitutively stable expressing cell # 10. Out of the total N- glycoside-linked sugar chain in which each cell is linked to a heavy chain constant region that is included in the antibody composition that has been purified including breaks IV anti-CAPRIN-1 antibody # 0 to # 10 to produce, sugar reduction the proportion of anti-caprin-1 antibodies at the ends of the N- acetylglucosamine having a sugar chain in which fucose is not bound LabChip (registered trademark) GXII was (PerkinElmer, Inc.) evaluation result both 80% or more.

[0249]

　Example 7: Modified anti-caprin-1 antibody of the antigen specificity and reactivity to cancer cells
　modified type I anti-caprin-1 antibody prepared in Examples 6-1 to 3 # 0 ~ # 10, modified type II anti CAPRIN-1 antibody # 0 to # 10, the modified type III anti-CAPRIN-1 antibody # 0 to # each antibody composition containing 10 and Aratameko type IV CAPRIN-1 antibody # 0 to # each antibody composition containing 10 ( the specific reactivity to caprin-1 protein in hereinafter referred to as modified anti-caprin-1 antibody) was confirmed in the same manner as in example 4. As a result, the absorbance values ​​of the wells supplemented with human IgG antibody does not react to caprin-1 protein used as negative controls have been tested, but were equally low values ​​and the well without addition of the antibody against, the absorbance values ​​of the wells was added the modified anti-caprin-1 antibodies, respectively showed all equally high. Moreover, all the modified anti-caprin-1 antibodies to the wells that do not solid phase the caprin-1 protein showed only negative control comparable absorbance values. From this result, all of the modified anti-caprin-1 antibody was confirmed to react specifically to caprin-1 protein.

[0250]

　Next, to confirm the reactivity to various human cancer cells in each modified anti-caprin-1 antibody in the same manner as in Example 4. Breast cancer cells (BT-474, MDA-MB-361), colon cancer cells (HT-29), lung cancer cells (QG56), gastric cancer cells (NCI-N87), uterine cancer cells (HEC-1-A), prostate cancer cell (22Rv1), pancreatic cancer cells (Panc10.5), liver cancer cells (Hep3B), ovarian cancer cells (SKOV3), renal carcinoma cells (Caki-2), brain tumor cells (U-87MG), bladder cancer cells (T24 , HT-1376), esophageal cancer cells (OE33), leukemia cells (OCI-AML5), lymphoma cells (Ramos), gallbladder cancer cells (TGBC14TKB), fibrosarcoma cells (HT-1080), melanoma cells (G-361) , adrenocortical carcinoma cells (A-673), Ewing's tumor cells (RD-ES), Hodgkin's lymphoma cells (RPMI1666), mesothelioma cells (NCI-H2452), multiple myeloma cells (IM-9), testicular cancer cells (NT / D1), thyroid cancer cells (TT), a head and neck cancer cells (FaDu) was used in this evaluation. As a result, in all of the cancer cells used in the evaluation, fluorescence intensity of each modified anti-caprin-1 antibody was stronger than the fluorescence intensity in the case of using the negative control. From the above results, all the modified anti-caprin-1 antibody, it was confirmed that react specifically to caprin-1 protein present on human cancer cell membrane surface.

[0251]

　Example 8: modified anti-CAPRIN-1 antibody of the various types of human cancer cells to anti-tumor activity
　in Example 6-1 was prepared in to 6-3 modified anti-CAPRIN-1 antibody (modified type I anti-CAPRIN-1 antibody # 0 ~ # 10, the modified type II anti CAPRIN-1 antibody # 0 to # 10, the modified type III anti-CAPRIN-1 antibody # 0 to # each antibody composition containing 10 and reforming IV anti-CAPRIN-1 antibody # 0 to # the anti-tumor effect against various human cancer cell of each antibody composition) containing 10 in the same manner as in example 5 were evaluated, respectively ADCC activity. As negative control, wells were added isotype control antibody reacts antibody to wells and caprin-1 protein without the addition of, were added to an antibody that is-reactive with human carcinoma cell surface caprin-1 is expressed It was prepared well. As a comparison antibody with the humanized antibody # 0 to # 10 are each anti-caprin-1 antibody prior to modification. Each antibody final concentration were added to V-bottom 96-well plate so that 0.01 ~ 1μg / mL.

[0252]

　Ha target cells, breast cancer cells (BT-474, MDA-MB -361), colon carcinoma (HT-29), lung cancer cells (QG56), gastric cancer cells (NCI-N87), uterine cancer (HEC-1-A ), before the legislature adenocarcinoma cells (22Rv1), pancreas Zang cancer (Panc10.5), liver Zang cancer cells (Hep3B), egg Chao cancer cells (SKOV3), renal cell carcinoma (Caki-2), Nao neoplasm cells (U-87MG), bladder cancer cells (T24, HT-1376), esophageal cancer cells (OE33), leukemia cells (OCI-AML5), ri nn pa swollen cells (Ramos), bile Nang cancer (TGBC14TKB), line-dimensional flesh swollen cells (HT-1080),メGetting Techno have ma cells ( G-361), adrenal cortical carcinoma (A-673), yuーイnn Corning neoplasm cells (RD-ES),ホji ki nn ri nn pa swollen cells (RPMI1666), the mesothelioma cells (NCI-H2452), multi Requested Procedure bone marrow swollen cells (IM-9 ), testicular cancer (NT / D1), thyroid cancer (TT), head and neck cancer (FaDu) with i ta wo. 10 . 6 remember Hikaru Suites cancer cell lines woそme cryぞme cry 50mL capacityのpreviousのtelecentric chiューbu ni set Circular, 100μCiのku ro ミウRousseau 51 (AiluropodaーDaikin e ru ma have Inc.) wo plusえ37 ℃で1 Time DIC CorporationュベーSuites shi after ta, 10%のFBS wo containingむRPMI1640 training groundで3 back wash shi, before the mind of each antibody wo shi ta add 96 points V bottom ni pu RitzーSuites, 1 hole thou ta ri × 10 2 3 Ge zu zu shi te add anti Applied sa se ​​ta.

[0253]

　Effector cells, using human NK cells isolated using conventional methods from human peripheral blood mononuclear cells. Above each antibody, the target cells with human NK cells in V-bottom 96-well plates that are carried out with the addition, 0.4 ~ 2.0 × 10 per well 5 Prepare those pieces added, 37 ° C., 5% , CO 2 was allowed to react for 4 hours under the conditions of. After the reaction, the culture supernatant containing the chromium 51 to be released from cancer cells that have undergone a failure to recover each well 50μL, on a culture that is released from cancer cells that have undergone a failure in the same manner as in Example 5 measuring the amount of chromium 51 in the supernatant was calculated antitumor effects on cancer cells by anti-caprin-1 antibody.

[0254]

　As a result, with respect to breast cancer cells (BT-474), a modified anti-CAPRIN-1 is an antibody modified type I anti-CAPRIN-1 antibody # 0 to # 10, the modified type II anti CAPRIN-1 antibody # 0 to # 10 , each antibody composition comprising a modified type III anti-CAPRIN-1 antibody # 0 each antibody compositions containing ~ # 10 and breaks IV anti-CAPRIN-1 antibody # 0 to # 10, strong anti respectively compared to the negative control It showed the tumor effect. In addition, the antibody prior to the modification of a comparative antibody (humanized antibody # 0 to # 10) breaks I type the same anti-tumor effect and anti-tumor effect indicated by the respective anti-CAPRIN-1 antibody # 0 to # 10, the modified type II antibody concentration upon illustrating each antibody composition comprising an anti-caprin-1 antibody # 0 to # 10 and reforming type III anti-caprin-1 antibody # 0 to # 10 is approximately as compared with the antibody concentration before modification 13 It was one of the concentration to 20 minutes. Further, if each antibody composition comprising a modified IV anti-caprin-1 antibody # 0 to # 10 to obtain the same anti-tumor effect as the antibody prior to modification is approximately as compared with the antibody concentration before modification 150 It was one of a concentration of min. As a result of the above from the modified anti-CAPRIN-1 antibody (modified type I anti-CAPRIN-1 antibody # 0 to # 10, the modified type II anti CAPRIN-1 antibody # 0 to # 10, the modified type III anti-CAPRIN-1 antibody # 0 each antibody composition comprising at ~ # 10 and modified IV anti-caprin-1 antibody # 0 ~ # 10), it was found that antitumor activity is enhanced as compared to the antibody prior to modification. Each antibody composition comprising a modified IV anti-caprin-1 antibody # 0 ~ # 10, modified I-type anti-caprin-1 antibody # 0 ~ # 10, modified type II anti-caprin-1 antibody # 0 ~ # 10 and it was found that a stronger anti-tumor effect compared to the respective antibody composition comprising a modified type III anti-caprin-1 antibody # 0 to # 10 is obtained.

[0255]

　Others, breast cancer cells that were used in the evaluation (MDA-MB-361), colon cancer cells (HT-29), lung cancer cells (QG56), gastric cancer cells (NCI-N87), uterine cancer cells (HEC-1-A), prostate cancer cells (22Rv1), pancreatic cancer cells (Panc10.5), liver cancer cells (Hep3B), ovarian cancer cells (SKOV3), renal carcinoma cells (Caki-2), brain tumor cells (U-87MG), bladder cancer cells (T24, HT-1376), esophageal cancer cells (OE33), leukemia cells (OCI-AML5), lymphoma cells (Ramos), gallbladder cancer cells (TGBC14TKB), fibrosarcoma cells (HT-1080), melanoma cells (G- 361), adrenal cortical cancer cells (A-673), Ewing's tumor cells (RD-ES), Hodgkin's lymphoma cells (RPMI1666), mesothelioma cells (NCI-H2452), multiple myeloma cells (IM-9), testicular cancer cells (NT / D1), thyroid cancer cells (TT), a strong anti-tumor effect was obtained by the same for head and neck cancer cells (FaDu).

Industrial applicability

[0256]

　Antibodies of the present invention are useful for the treatment and / or prevention of cancer.

[0257]

　In addition, all of the publications cited in this specification, it is assumed that incorporate herein by patents and patent applications as it is as a reference.

The scope of the claims

[Claim 1]

　And a light chain variable region comprising the heavy chain complementarity determining regions of the variable region to SEQ ID NO: 4, 5 and 6 including the complementarity determining regions of SEQ ID NO: 1, 2 and 3, and, caprin-1 protein and immunologically antibody or fragment thereof having reactive.

[Claim 2]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 8, and antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 15.

[Claim 3]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 10, and an antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 15.

[Claim 4]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 7 and an antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 15.

[Claim 5]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 8, and antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 13.

[6.]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 7 and an antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 12.

[7.]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 8, and antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 12.

[8.]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 7 and an antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 13.

[9.]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 10, and an antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 14.

[10.]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 8, and antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 11.

[11.]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 7 and an antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 11.

[12.]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 9 and an antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 15.

[13.]

　It comprises the amino acid sequence of the heavy chain variable region SEQ ID NO: 20, and an antibody or fragment thereof according to claim 1 light chain variable region comprising the amino acid sequence of SEQ ID NO: 21.

[14.]

　Human antibodies, humanized antibodies, chimeric antibodies, a single chain antibody or a multispecific antibody, antibody or fragment thereof according to any one of claims 1 to 13.

[15.]

　Antibody or fragment thereof according to any one of claims 1 to 13, anti-tumor agent conjugated.

[16.]

　Wherein including one or more amino acid substitutions in the heavy chain constant region of the antibody, the antibody according to any one of claims 1 to 15.

[17.]

　Wherein said antibody is an antibody obtained by removing fucose bound to N- acetylglucosamine sugar chain reducing end of N- glycoside-linked sugar chain which binds to the heavy chain constant region, according to any one of claims 1 to 16 antibody of.

[18.]

　The composition of the antibody, according to claim 17 claims 1-16 which fucose is bound to the antibody and heavy chain binds to the constant region of the N- glycoside-linked sugar chain carbohydrate reducing end of N- acetylglucosamine as described in comprising the antibody according to item 1 in any of the antibody of the composition.

[19.]

　Cells producing a composition of an antibody according to an antibody or claim 18 of claim 17.

[20.]

　Antibody or fragment thereof according to any one of claims 1 to 15, characterized in that it comprises antibodies according to claim 16 or 17, or a composition of an antibody according to claim 18 as an active ingredient, the pharmaceutical compositions for the treatment and / or prevention of cancer.

[21.]

　Wherein the cancer is breast cancer, renal cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer, stomach cancer, uterine cancer, ovarian cancer, prostate cancer, bladder cancer, esophageal cancer, leukemia, lymphoma, liver cancer, gallbladder cancer, sarcoma, mastocytoma , melanoma, adrenocortical cancer, Ewing's tumor, Hodgkin's lymphoma, mesothelioma, multiple myeloma, testicular cancer, thyroid cancer, or head and neck cancer, the pharmaceutical composition according to claim 20.

[22.]

　And claim 20 or a pharmaceutical composition according to 21, comprising a pharmaceutical composition comprising an anti-tumor agent, a combination medicament for the treatment and / or prevention of cancer.

[23.]

　DNA encoding the antibody or fragment thereof according to any one of claims 1 to 16.

[24.]

　Antibody or fragment thereof according to any one of claims 1 to 16, the antibody of claim 17, the composition of the antibody according to claim 18, the pharmaceutical composition according to claim 20 or 21, or the combination medicament according to claim 22, comprising administering to a subject, the treatment and / or prevention of cancer.