The disclosure relates generally to biology and medicine, and more particularly it relates to antibodies (Abs) that bind to and thereby neutralize human angiopoietin-like protein (ANGPTL) 3/8 complexes. Such Abs can increase lipoprotein lipase (LPL) activity and thereby lower serum triglycerides (TGs) such that they may be used in treating lipid metabolism-related and glucose metabolism-related diseases and disorders.

[0002] ANGPTLs are a family of proteins that regulate a number of physiological and pathophysiological processes. Of particular interest herein, is the role of ANGPTL3 and ANGPTL8 in lipid and glucose metabolism.

[0003] Evidence supports the role of ANGPTL3 as a main regulator of lipoprotein metabolism, and it may regulate TG clearance by inhibiting LPL and inhibiting endothelial lipase (EL). See , Chi et al. (2017) Mol. Metab. 6: 1137-1149. ANGPTL3 deficiency, inactivation, or loss can result in low levels of low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C) and TGs. ANGPTL3 also may affect insulin sensitivity, thereby playing a role in modulating not only lipid metabolism but also glucose metabolism. See, Robciuc etal. (2013) Arterioscler. Thromb. Vase. Biol. 33: 1706-1713. Nucleic acid and amino acid sequences for human ANGPTL3 are known. For example, one nucleic acid sequence can be found in NCBI Reference Sequence No. NM 014495 (SEQ ID NO: l), and one amino acid sequence can be found in NCBI Reference Sequence No. NP_055310 (SEQ ID NO:2).

[0004] ANGPTL8 is highly expressed in the liver and adipose tissue and has been reported to inhibit LPL by complexing with and thereby activating ANGPTL3. See , Chi, supra. Human ANGPTL8 appears to be induced by feeding. Nucleic acid and amino acid sequences for human ANGPTL8 are known. For example, one nucleic acid sequence can be found in NCBI Reference Sequence No. NM 018687 (SEQ ID NO:3), and one amino acid sequence can be found in NCBI Reference Sequence No. NP 061157 (SEQ ID NO:4).

[0005] ANGPTL3/8 complexes exist, which have one or more ANGPTL3s that are bound to one or more ANGPTL8s. Evidence suggests these complexes more effectively mediate inhibition of LPL when compared to ANGPTL3 or ANGPTL8 alone. Moreover,

ANGPTL3/8 complexes may be made in vitro by co-expressing ANGPTL8 and ANGPTL3 in a mammalian expression system. See , Chi, supra.

[0006] Abs are known that bind to either ANGPTL3 or ANGPTL8 and that can be used alone or in combination with each other to treat lipid metabolism-related and glucose metabolism-related diseases and disorders. For example, Inti. Patent Application Publication No. WO 2012/174178 discloses a fully human monoclonal Ab and antigen binding fragments thereof that bind to ANGPTL3 and interfere with its activity. Other therapeutic anti-ANGPTL3 Abs also are known. See, e.g. , Inti. Patent Application Publication No. WO 2008/073300 and US Patent No. 7,935,796. Likewise, Inti. Patent Application Publication No. WO 2017/027316 discloses a fully human monoclonal Ab or antigen-binding fragments thereof that bind to ANGPTL8 and interfere with its activity. Moreover, Inti. Patent Application Publication No. WO 2017/177181 discloses a combined anti-ANGPTL3 Ab and anti-ANGPTL8 Ab therapy.

[0007] Unfortunately, existing Abs that bind only to either ANGPTL3 or ANGPTL8 do not fully abrogate the effect of these ANGPTLs and/or ANGPTL3/8 complexes on lipid and/or glucose metabolism. See, e.g., Dewey el al. (2017) N. Engl. J. Med. 377:211-221; and Gusarova et al. (2017) Endocrinology 158: 1252-1259. In view thereof, there is a need for additional Abs, especially anti-ANGPTL3/8 complex Abs, for treating lipid metabolism-related and glucose metabolism-related diseases and disorders, where such Abs have improved pharmacological inhibitory and/or regulatory properties to modulate lipid and/or glucose metabolism.

[0008] To address this need, nucleic and amino acid sequences are provided for a modified ANGTPL3/8 complex. Accordingly, nucleic acid sequences encoding one or more of a modified ANGPTL3 and a modified ANGPTL8 (i.e., fusion proteins) are described herein. In some instances, the nucleic acid sequences include a polynucleotide sequence encoding an ANGPTL3 fusion protein having an amino acid sequence of SEQ ID NO: 17. In other instances, the nucleic acid sequences include a polynucleotide sequence encoding an ANGPTL8 fusion protein having an amino acid sequence of SEQ ID NO: 18. In still other instances, the nucleic acid sequences include a polynucleotide sequence encoding SEQ ID NO: 17 and 18.

[0009] Additionally, nucleic acid constructs are provided that include a polynucleotide sequence encoding an ANGPTL3 fusion protein as described herein, an ANGPTL8 fusion

protein as described herein, or both, where such constructs can be an expression cassette or a vector.

[0010] In view of the above, host cells are provided that include therein one or more expression cassettes or vectors as described herein. In some instances, the host cells are eukaryotic cells. In some instances, the polynucleotide sequences for the ANGPTL3 and ANGTPL8 fusion proteins are on separate expression cassettes or vectors, while in other instances they can be on the same expression cassette or vector.

[0011 ] Also, ANGPTL3 fusion proteins are provided that include an amino acid sequence of SEQ ID NO: 17 or 19, as well as active variants or fragments thereof. Likewise, ANGPTL8 fusion proteins are provided that include an amino acid sequence of SEQ ID NO: 18 or 20, as well as active variants or fragments thereof.

[0012] Moreover, functional ANGPTL3/8 complexes are provided, especially human ANGPTL3/8 complexes, where an ANGPTL3 moiety of the complex is a native (full-length or truncated) ANGPTL3 or an ANGPTL3 fusion protein as described herein and where an ANGPTL8 moiety of the complex is an ANGPTL8 fusion protein as described herein. In some instances, the ANGPTL3 fusion protein includes an amino acid sequence of SEQ ID NO: 19. Likewise, and in some instances, the ANGPTL8 fusion protein includes an amino acid sequence of SEQ ID NO:20. Moreover, and in some instances, the complexes can have a 1 : 1 ratio of the ANGPTL3 moiety to the ANGPTL8 moiety. In other instances, the complexes can have ratios other than 1 : 1, such as 1 :2, 1 :3, 2: 1 or 3 : 1 ratio of ANGPTL3 moiety to ANGPTL8 moiety, respectively.

[0013] Methods also are provided for making recombinant ANGTPL3/8 complexes. The methods can include at least a step of expressing one or more polynucleotide sequences for an ANGPTL3 moiety and an ANGPTL8 moiety as described herein in a host cell such as in a mammalian expression system to obtain ANGPTL3/8 complexes therefrom. In some instances, the ANGPTL3 and ANGPTL8 moieties are provided on separate expression constructs or vectors. In other instances, ANGPTL3 and ANGPTL8 are provided on one expression construct or vector. The methods also can include a step of purifying the resulting ANGPTL3/8 complexes, which may include not only concentrating the ANGPTL3/8 complexes but also removing one or more of the tags, linkers and serum albumin from the ANGPTL3 moiety and/or the ANGPTL8 moiety. The methods also can

include a step of concentrating the ANGPTL3/8 complexes before and/or after purifying step.

[0014] Second, an Ab to a ANGPTL3/8 complex is provided as well as uses thereof, which includes treating lipid metabolism-related and glucose metabolism-related diseases and disorders by binding to and thereby inhibiting ANGPTL3/8 complex activity.

[0015] An effective amount of the anti-ANGPTL3/8 complex Ab described herein, or a pharmaceutically acceptable salt thereof, may be used for increasing LPL activity, lowering TGs, and treating lipid metabolism- and/or glucose metabolism-related diseases or disorders in an individual in need thereof.

[0016] The anti-ANGPTL3/8 complex Ab described herein binds soluble ANGPTL3/8 complex, thereby increasing LPL activity and decreasing serum TG levels. Individuals with lower TG levels are at lower risk for cardiovascular disease. Advantageously, the anti-ANGPTL3/8 complex Ab described herein binds only to the ANGPTL3/8 complex and not to ANGPTL3 alone or to ANGPTL8 alone at relevant concentrations. It is believed that the anti-ANGPTL3/8 complex Ab increases catabolism of TG-rich lipoproteins (TRLs), which reduces TGs and/or non-HDL-C, thereby improving dyslipidemia risk factors for atherosclerotic cardiovascular disease (ASCVD) not addressed by current therapies. Moreover, and because the anti-ANGPTL3/8 complex Ab described herein does not bind ANGPTL3 or ANGPTL8 alone, other actions of these ANGPTLs are not inhibited, which can lead to fewer untoward in vivo effects, such as reduced de-repression of EL.

[0017] In particular, the anti-ANGPTL3/8 complex Ab is a human anti-ANGPTL3/8 complex Ab. In some instances, the anti-ANGPTL3/8 complex Ab can abrogate, block, inhibit, interfere, neutralize or reduce in vivo activity of the ANGPTL3/8 complex, especially its LPL inhibitory activity. In some instances, the anti-ANGPTL3/8 complex Ab can be full-length or can be only an antigen-binding fragment ( e.g ., a Fab, F(ab’)2 or scFv fragment). Desirable properties of an anti-ANGPTL3/8 complex Ab include TG lowering at low doses of the Ab, that is durable for at least 21 days.

[0018] In some instances, the anti-ANGPTL3/8 complex Ab binds human ANGPTL3/8 complex and includes light chain determining regions LCDR1, LCDR2 and LCDR3 and heavy chain determining regions HCDR1, HCDR2 and HCDR3, where LCDR1 has the amino acid sequence RSSQSLLDSDDGNTYLD (SEQ ID NO: 11), LCDR2 has the amino acid sequence YMLSYRAS (SEQ ID NO: 12) and LCDR3 has the amino acid sequence

MQRIEFPLT (SEQ ID NO: 13), and where HCDR1 has the amino acid sequence TFSGFSLSISGVGVG (SEQ ID NO: 14), HCDR2 has the amino acid sequence LIYRNDDKRY SP SLK S (SEQ ID NO: 15) and HCDR3 has the amino acid sequence ARTY S S GW Y GNWFDP (SEQ ID NO: 16).

[0019] Further provided is an Ab including a light chain variable region (LCVR), where the LCVR has the amino acid sequence of SEQ ID NO: 9; or an Ab including a heavy chain variable region (HCVR), where the HCVR has the amino acid sequence of SEQ ID NO: 10. In some instances, the Ab includes an LCVR with the amino acid sequence of SEQ ID NO:9 and a HCVR with the amino acid sequence of SEQ ID NO: 10. In some instances, the Ab includes a light chain (LC) and a heavy chain (HC), where the LC has the amino acid sequence of SEQ ID NO:5 or the HC has the amino acid sequence of SEQ ID NO:6. Alternatively, the Ab includes a light chain (LC) and a heavy chain (HC), where the LC has the amino acid sequence of SEQ ID NO: 5 and the HC has the amino acid sequence of SEQ ID NO:6. In certain instances, the Ab is an IgG4 isotype.

[0020] In some instances, the anti-ANGPTL3/8 complex Ab can be a variant of the Ab described above, especially a LC variant having a D31 S mutation (SEQ ID NO:21), a D33 A mutation (SEQ ID NO:22), a D33T mutation (SEQ ID NO:23); a M56T mutation (SEQ ID NO:24), a E99Q mutation (SEQ ID NO:25) or a combination thereof (e.g, D33T and M56T mutations or D33A and M56T mutations), with respect to a LC having an amino acid sequence of SEQ ID NO: 5.

[0021] Furthermore, an Ab is provided that is produced by cultivating a mammalian cell including a cDNA molecule, where the cDNA molecule encodes polypeptides having the amino acid sequences of SEQ ID NO: 5 and 6, under such conditions that the polypeptides are expressed, and recovering the Ab. Alternatively, an Ab is provided that is produced by cultivating a mammalian cell including two cDNA molecules, where a first cDNA molecule encodes a polypeptide having the amino acid sequence of SEQ ID NO:5, and a second cDNA molecule encodes a polypeptide having the amino acid sequence of SEQ ID NO: 6, under such conditions that the polypeptides are expressed, and recovering the Ab. In some instances, the anti-ANGPTL3/8 complex Ab can be a variant of the Ab described above, especially a LC variant having a D31 S mutation (SEQ ID NO:21), a D33 A mutation (SEQ ID NO: 22), a D33T mutation (SEQ ID NO:23); a M56T mutation (SEQ ID NO:24), a E99Q

mutation (SEQ ID NO:25) or a combination thereof, with respect to a LC having an amino acid sequence of SEQ ID NO: 5.

[0022] Moreover, an Ab is provided that binds to and neutralizes human ANGPTL3/8 complex in a standard LPL activity assay with an EC50 of 0.5 nM or less. Also, an Ab is provided that binds to human ANGPTL3/8 complex with a dissociation constant of less than or equal to 1 x 10 6 M. Moreover, an Ab is provided that binds to human ANGPTL3 and human ANGPTL8 with a dissociation constant of greater than 1 x 10 6 M. Furthermore, an Ab is provided that binds to human ANGPTL3/8 complex with a signal greater than 3 fold over the non-binding background signal, as measured by single point ELISA assay, but does not bind to human ANGPTL3 alone or human ANGPTL8 alone with a signal greater than 3 fold over the non-binding background signal, as measured by single point ELISA assay. Likewise, an Ab is provided that lowers TGs in vivo by at least 50% when compared to IgG control at a dose of 10 mg/kg at a time point 14 days after dosing.

[0023] Third, a pharmaceutical composition is provided that includes the Ab herein or a population of Abs herein and an acceptable carrier, diluent or excipient. Also provided is a mammalian cell including a DNA molecule including a polynucleotide sequence encoding polypeptides having amino acid sequences of SEQ ID NO: 5 and SEQ ID NO: 6, where the cell is capable of expressing an Ab herein. Further provided is a mammalian cell including a first DNA molecule and a second DNA molecule, where the first DNA molecule includes a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO:5, and where the second DNA molecule includes a polynucleotide sequence encoding a polypeptide having the amino acid sequence of SEQ ID NO: 6, and where the cell is capable of expressing an Ab herein. In some instances, the anti-ANGPTL3/8 complex Ab can be a variant of the Ab described above, especially a LC variant having a D31 S mutation (SEQ ID NO:21), a D33A mutation (SEQ ID NO:22), a D33T mutation (SEQ ID NO:23); a M56T mutation (SEQ ID NO:24), a E99Q mutation (SEQ ID NO:25) or a combination thereof, with respect to a LC having an amino acid sequence of SEQ ID NO: 5.

[0024] Fourth, a process is provided for producing an Ab, where the process includes cultivating a mammalian cell with a DNA molecule having a polynucleotide sequence encoding polypeptides having the amino acid sequences of SEQ ID NO: 5 and SEQ ID NO:6, where the cell is capable of expressing the Ab herein under conditions such that the Ab is expressed, and recovering the expressed Ab. In some instances, the polynucleotide sequence encoding the polypeptide having the amino acid sequence of SEQ ID NO: 5 can encode a D31S mutation, a D33A mutation, a D33T mutation, a M56T mutation, a E99Q mutation or a combination thereof. Also provided herein is a method of treating ASCVD, chronic kidney disease (CKD), diabetes, hypertriglyceridemia, nonalcoholic steatohepatitis (NASH), obesity, or a combination thereof, where the method includes administering to an individual in need thereof, an effective amount of an Ab herein. Further provided is a method of lowering TGs that includes administering to an individual in need thereof, an effective amount of an Ab herein.

[0025] Fifth, an Ab is provided for use in therapy. In particular, the Ab is for use in the treatment of ASCVD, CKD, diabetes, hypertriglyceridemia, NASH, obsesity, or a combination thereof. Also provided is a pharmaceutical composition for use in treating ASCVD, CKD, diabetes, hypertriglyceridemia, NASH, obsesity, or a combination thereof, that includes an effective amount of the an the Ab herein.

[0026] The advantages, effects, features and objects other than those set forth above will become more readily apparent when consideration is given to the detailed description below. Such detailed description makes reference to the following drawing(s), where:

[0027] FIG. 1 shows an image of a SDS-page gel showing ANGPTL3/8 complex run under reduced and non-reduced conditions.

[0028] Reference to an element by the indefinite article“a” or“an” does not exclude the possibility that more than one element is present, unless the context clearly requires that there be one and only one element. The indefinite article“a” or“an” thus usually means “at least one.”

[0029] Definitions

[0030] As used herein,“about” means within a statistically meaningful range of a value or values such as, for example, a stated concentration, length, molecular weight, pH, sequence similarity, time frame, temperature, volume, etc. Such a value or range can be within an order of magnitude typically within 20%, more typically within 10%, and even more typically within 5% of a given value or range. The allowable variation encompassed by“about” will depend upon the particular system under study, and can be readily appreciated by one of skill in the art.

[0031] As used herein,“affinity” means a strength of an Ab’s binding to an epitope on an ANGPTL3/8 complex.

[0032] As used herein,“angiopoietin-like protein 3” or“ANGPTL3” means a protein having an amino acid sequence including SEQ ID NO:2.

[0033] As used herein,“angiopoietin-like protein 8” or“ANGPTL8” means a protein having an amino acid sequence including SEQ ID NO:4.

[0034] As used herein,“ANGPTL3/8 complex” means a multi-protein complex of one or more ANGPTL3 compounds that are bound to one or more ANGPTL8 compounds.

[0035] As used herein,“anti-ANGPTL3/8 complex Ab” or“anti-ANGPTL3/8 complex Ab” means an Ab that simultaneously recognizes and binds to an area on both ANGPTL3 and ANGPTL8, especially when in the form of the ANGPTL3/8 complex. Generally, an anti-ANGPTL3/8 complex Ab will usually not bind to other ANGPTL family members (e.g., ANGPTL 1, ANGPTL2, ANGPTL4, ANGPTL5, ANGPTL6 or ANGPTL7). Moreover, and as noted elsewhere, an anti-ANGPTL3/8 complex Ab also will not bind to ANGPTL3 or ANGPTL8 alone at specified concentrations, as described in the single point ELISA assay below.

[0036] As used herein,“bind” or“binds” means an ability of a protein to form a type of chemical bond or attractive force with another protein or molecule as determined by common methods known in the art. Binding can be characterized by an equilibrium dissociation constant (KD) of about 1 xlO 6 M or less (i.e., a smaller KD denotes a tighter binding). Methods of determining whether two molecules bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. Here, an anti-ANGPTL3/8 complex Ab binds only the ANGPTL3/8 complex and does not bind ANGPTL3 alone or ANGPTL8 alone. Whether an Ab binds only to the ANGPTL3/8 complex and not to ANGPTL3 alone or ANGPTL8 alone can be determined in standard ELISA assays in a single point format, as described below and binding may be characterized by Biacore, as described below. While the Abs herein are human, they may, however, exhibit cross-reactivity to other ANGPTL3/8 complexes from other species, for example, cynomolgus monkey ANGPTL3/8 complex, mouse ANGPTL3/8 complex, or rat ANGPTL3/8 complex.

[0037] As used herein,“effective amount” means an amount or dose of a compound or a pharmaceutical composition containing the same, which upon single or multiple dose

administration to an individual, will elicit a biological or medical response of or desired therapeutic effect on a tissue, system, animal, mammal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician. In some instances, an effective amount of a compound herein or compositions including the same to an individual in need thereof would result in increasing LPL activity. A dose can include a higher initial loading dose, followed by a lower dose. A dose can be administered in any therapeutically effective interval, such as multiple times a day, once daily, every other day, three times a week, two times a week, one time a week, once every two weeks, once a month, once every two months, etc. A dose constituting an effective amount could be between 0.01 mg/kg and 100 mg/kg.

[0038] As used herein,“equilibrium dissociation constant” or“KD” means a quantitative measurement of Ab affinity to a particular antigen interaction, such as affinity of an Ab to ANGPTL3/8 complex, especially a measure of a propensity of an Ab/ANGPTL3/8 complex conjugate to separate reversibly into its component parts. Likewise, and as used herein,“equilibrium association constant” or“Ka” means an inverse of KD.

[0039] As used herein, “functional” means that and ANGPTL3 fusion protein, ANGPTL8 fusion protein or ANGPTL3/8 complex has biological activity akin to that of a native ANGPTL3, a native ANGPTL8 or a native ANGPTL3/8 complex including, for example, inhibiting LPL or acting as an antigen to which an Ab can be made and directed.

[0040] As used herein,“glucose metabolism-related disease or disorder” means diabetes and the like.

[0041] As used herein,“lipid metabolism-related disease or disorder” means a condition associated with abnormal lipid metabolism such as dyslipidemia, hyperlipidemia and hyperlipoproteinemia, including hypertriglyceridemia, hypercholesterolemia, chylomicronemia, mixed dyslipidemia (obesity, metabolic syndrome, diabetes, etc.), lipodystrophy and lipoatrophy. The term also encompasses certain cardiovascular diseases such as atherosclerosis and coronary artery disease, acute pancreatitis, NASH, obesity and the like.

[0042] As used herein, “half-maximal effective concentration” or“EC50” means a concentration of an Ab (typically expressed in molar units (M)) that induces a response halfway between a baseline and a maximum after a predetermined period of time. The EC50 described herein is ideally 3.0 nM or less.

[0043] As used herein,“nucleic acid construct” or“expression cassette” means a nucleic acid molecule having at least a control sequence operably linked to a coding sequence. In this manner, a control sequence such as a promoter is in operable interaction with nucleic acid sequences encoding at least one polypeptide of interest such as the ANGPTL3 fusion proteins described herein and/or the ANGPTL8 fusion proteins described herein. Such nucleic acid constructs can be in the form of an expression or transfer cassette. A nucleic acid construct can include an oligonucleotide or polynucleotide composed of deoxyribonucleotides, ribonucleotides or combinations thereof having incorporated therein the nucleotide sequences for one or more polypeptides of interest.

[0044] As used herein,“operably linked” means that the elements of a nucleic acid construct are configured so as to perform their usual function. Thus, control sequences (i.e., promoters) operably linked to a coding sequence are capable of effecting expression of the coding sequence. The control sequences need not be contiguous with the coding sequence, so long as they function to direct the expression thereof (i.e., maintain proper reading frame). Thus, for example, intervening untranslated, yet transcribed sequences, can be present between a promoter and a coding sequence, and the promoter sequence still can be considered“operably linked” to the coding sequence.

[0045] As used herein,“control sequence” or“control sequences” means promoters, polyadenylation signals, transcription and translation termination sequences, upstream regulatory domains, origins of replication, internal ribosome entry sites (“IRES”), enhancers, and the like, which collectively provide for replication, transcription and translation of a coding sequence in a recipient host cell. Not all of these control sequences need always be present so long as the selected coding sequence is capable of being replicated, transcribed and translated in an appropriate host cell.

[0046] As used herein,“coding sequence” or“coding sequences” means a nucleic acid sequence that encodes for one or more polypeptides of interest, and is a nucleic acid sequence that is transcribed (in the case of DNA) and translated (in the case of RNA) into a polypeptide in vitro or in vivo when placed under the control of appropriate regulatory sequences. The boundaries of the coding sequence(s) are determined by a start codon at a 5’ (amino) terminus and a translation stop codon at a 3’ (carboxy) terminus. A coding sequence can include, but is not limited to, viral nucleic acid sequences, cDNA from

prokaryotic or eukaryotic mRNA, genomic DNA sequences from prokaryotic or eukaryotic DNA, or even synthetic DNA sequences.

[0047] With respect to control and coding sequences, they can be native/analogous to the host cell or each other. Alternatively, the control and coding sequences can be heterologous to host cell or each other.

[0048] As used herein,“promoter” means a nucleotide region composed of a nucleic acid regulatory sequence, where the regulatory sequence is derived from a gene or synthetically created and is capable of binding RNA polymerase and initiating transcription of a downstream (3’ -directi on) coding sequence. A number of promoters can be used in nucleic acid constructs, including a native promoter for one or more polypeptides of interest. Alternatively, promoters can be selected based upon a desired outcome. Such promoters can include, but are not limited to, inducible promoters, repressible promoters and constitutive promoters.

[0049] As used herein,“variant” means a polynucleotide or a polypeptide having one or more modifications such as an addition, deletion, insertion and/or substitution of one or more specific nucleic acid or amino acid residues when compared to a reference nucleic acid or amino acid sequence. A variant therefore includes one or more alterations when compared to the reference nucleic acid or amino acid sequence. Here, the anti-ANGPTL3/8 complex Ab can have a LC or HC variation. In particular, the Ab can be a LC variant having a D31 S mutation (SEQ ID NO:21), a D33A mutation (SEQ ID NO:22), a D33T mutation (SEQ ID NO:23); a M56T mutation (SEQ ID NO:24) or a E99Q mutation (SEQ ID NO:25) with respect to a LC having an amino acid sequence of SEQ ID NO: 5. Likewise, the LC variant can be a combination any two of the above such as, for example, D31 S and D33 A mutations, D31 S and D33T mutations, D31 S and M56T mutations, D31 S and E99Q mutations, D33A and M56T mutations, D33A and E99Q mutations, D33T and M56T mutations, D33T and E99Q mutations, and M56T and E99Q, again with respect to a LC having an amino acid sequence of SEQ ID NO: 5. Moreover, the LC variant can be a combination of any three of the above such as, for example, D31 S, D33A and M56T mutations, D31 S, D33A and E99Q mutations, D31 S, D33T and M56T mutations, D31 S, D33T and E99Q mutaitons, D33A, M56T and E99Q mutations, and D33T, M56T and E99Q mutations, again with respect to a LC having an amino acid sequence of SEQ ID NO: 5. Furthermore, the LC variant can be a combination of any four of the above such as, for example, D31 S, D33 A, M56T and E99Q mutations; and D31 S, D33T, M56T and E99Q mutations, again with respect to a LC having an amino acid sequence of SEQ ID NO: 5.

[0050] As used herein,“vector” means a replicon, such as a plasmid, phage or cosmid, to which another nucleic acid sequence, such as an expression cassette, may be attached so as to bring about replication of the attached sequence. A vector is capable of transferring nucleic acid molecules to host cells. Vectors typically include one or a small number of restriction endonuclease recognition sites where a nucleotide sequence of interest can be inserted in a determinable fashion without loss of essential biological function of the vector, as well as a selectable marker that can be used for identifying and selecting host cells transformed with the vector. A vector therefor can be capable of transferring nucleic acid sequences to target cells.

CLAIMS

The invention claimed is:

1. A polynucleotide comprising a nucleic acid sequence encoding a polypeptide of SEQ ID NO: 17 or a polypeptide of SEQ ID NO: 18.

2. A polynucleotide encoding a nucleic acid sequence encoding a polypeptide of SEQ ID NO: 17 and a polypeptide of SEQ ID NO: 18.

3. An expression cassette comprising a polynucleotide of Claim 1 or 2.

4. A vector comprising a polynucleotide of Claim 1 or Claim 2.

5. A host cell comprising the expression construct of Claim 3 or the vector of Claim 4.

6. The host cell of Claim 5, wherein the host cell is a mammalian cell.

7. A polypeptide comprising an amino acid sequence of SEQ ID NO: 17 or 19.

8. A polypeptide comprising an amino acid sequence of SEQ ID NO: 18 or 20.

9. An angiopoietin-like peptide (ANGPTL)3/8 complex comprising a polypeptide having an amino acid sequence of SEQ ID NO: 19 and a polypeptide having an amino acid sequence of SEQ ID NO:20.