We claim:
1. An antibody-like binding protein comprising two polypeptide chains that form two
antigen binding sites, wherein a first polypeptide chain comprises the structure V1-L1-V2-L2-CL, and a second polypeptide chain comprises the structure V3-L3-V4-L4-CH1; wherein:
(a) V1 is a first immunoglobulin heavy chain variable domain (VH1), V2 is a
second immunoglobulin light chain variable domain (VL2), V3 is a second
immunoglobulin heavy chain variable domain (VH2), and V4 is a first immunoglobulin
heavy chain variable domain (VL1); or
V1 is a first immunoglobulin light chain variable domain (VL1), V2 is a second immunoglobulin heavy chain variable domain (VH2), V3 is a second immunoglobulin light chain variable domain (VL2), and V4 is a first immunoglobulin heavy chain variable domain (VH1); or
V1 is a first immunoglobulin heavy chain variable domain (VH1), V2 is a second immunoglobulin heavy chain variable domain (VH2), V3 is a second immunoglobulin light chain variable domain (VL2), and V4 is a first immunoglobulin light chain variable domain (VL1);
wherein VL1 and VH1 associate to form a first antigen binding site, and VL2 and VH2 associate to form a second antigen binding site;
(b) CL is an immunoglobulin light chain constant domain;
(c) CH1 is the immunoglobulin CH1 heavy chain constant domain;
(d) L1, L2, L3, and L4 are amino acid linkers;
(e) at least one of the first or second polypeptide chains further comprises a Fc domain; and
wherein the first and second polypeptides form a cross-over light chain-heavy chain pair.
2. The antibody-like binding protein of claim 1, wherein the binding protein is capable of specifically binding one or more antigen targets.
3. The antibody-like binding protein of claim 2, wherein the one or more antigen targets is selected from the group consisting of B7.1, B7.2, BAFF, BlyS, C3, C5, CCL11 (eotaxin), CCL15 (MIP-1d), CCL17 (TARC), CCL19 (MIP-3b), CCL2 (MCP-1), CCL20 (MIP-3a), CCL21 (MIP-2), SLC, CCL24 (MPIF-2/eotaxin-2), CCL25 (TECK), CCL26 (eotaxin-3),
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CCL3 (MIP-1a), CCL4 (MIP-1b), CCL5 (RANTES), CCL7 (MCP-3), CCL8 (mcp-2), CD3, CD19, CD20, CD24, CD40, CD40L, CD80, CD86, CDH1 (E-cadherin), Chitinase, CSF1 (M-CSF), CSF2 (GM-CSF), CSF3 (GCSF), CTLA4, CX3CL1 (SCYD1), CXCL12 (SDF1), CXCL13, EGFR, FCER1A, FCER2, HER2, IGF1R, IL-1, IL-12, IL13, IL15, IL17, IL18, IL1A, IL1B, IL1F10, IL1β, IL2, IL4, IL6, IL7, IL8, IL9, IL12/23, IL22, IL23, IL25, IL27, IL35, ITGB4 (b 4 integrin), LEP (leptin), MHC class II, TLR2, TLR4, TLR5, TNF, TNFα, TNFSF4 (OX40 ligand), TNFSF5 (CD40 ligand), Toll-like receptors, TREM1, TSLP, TWEAK, XCR1 (GPR5/CCXCR1), DNGR-1(CLEC91), and HMGB1.
4. The antibody-like binding protein of claim 1, wherein the binding protein is bispecific and capable of binding two different antigen targets.
5. The antibody-like binding protein of claim 4, wherein the two different antigen targets are selected from the group consisting of IL4 and IL13, IGF1R and HER2, IGF1R and EGFR, EGFR and HER2, BK and IL13, PDL-1 and CTLA-4, CTLA4 and MHC class II, IL-12 and IL-18, IL-1α and IL-1β, TNFα and IL12/23, TNFα and IL-12p40, TNFα and IL1β, TNFα and IL-23, and IL17 and IL23.
6. The antibody-like binding protein of claim 1, wherein the binding protein is capable of inhibiting the function of one or more of the antigen targets.
7. The antibody-like binding protein of claim 1, wherein at least one of the linkers selected from the group consisting of L1, L2, L3, and L4 contains at least one cysteine residue.
8. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of the antibody-like binding protein of claim 1.
9. The antibody-like binding protein of claim 1, wherein:
L1 is 3 to 12 amino acid residues in length;
L2 is 3 to 14 amino acid residues in length; L3 is 1 to 8 amino acid residues in length; and L4 is 1 to 3 amino acid residues in length.
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10. The antibody-like binding protein of claim 1, wherein:
L1 is 1 to 3 amino acid residues in length;
L2 is 1 to 4 amino acid residues in length;
L3 is 2 to 15 amino acid residues in length; and
L4 is 2 to 15 amino acid residues in length.
11. The antibody-like binding protein of claim 1, wherein:
L1 is 5 to 10 amino acid residues in length;
L2 is 5 to 8 amino acid residues in length; L3 is 1 to 5 amino acid residues in length; and L4 is 1 to 2 amino acid residues in length.
12. The antibody-like binding protein of claim 1, wherein:
L1 is 7 amino acid residues in length;
L2 is 5 amino acid residues in length; L3 is 1 amino acid residue in length; and L4 is 2 amino acid residues in length.
13. The antibody-like binding protein of claim 1, wherein:
L1 is 1 to 2 amino acid residues in length;
L2 is 1 to 2 amino acid residues in length;
L3 is 4 to 12 amino acid residues in length; and
L4 is 2 to 12 amino acid residues in length.
14. The antibody-like binding protein of claim 1, wherein:
L1 is 1 amino acid residue in length;
L2 is 2 amino acid residues in length; L3 is 7 amino acid residues in length; and L4 is 5 amino acid residues in length.
15. The antibody-like binding protein of claim 1, wherein the Fc domain is linked to the
carboxyl terminal region of the polypeptide chain.
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16. The antibody-like binding protein of claim 1, wherein the Fc domain is linked to CL and/or CH1.
17. The antibody-like binding protein of claim 14, wherein the Fc domain of the first polypeptide is linked to CL and/or the Fc domain of the second polypeptide is linked to the CH1.
18. An isolated nucleic acid molecule comprising a nucleotide sequence encoding the antibody-like binding protein of any one of claims 1-7 or 9-17.
19. An expression vector comprising the nucleic acid molecule of claim 18.
20. An isolated host cell comprising the nucleic acid molecule of claim 18 or the expression vector of claim 19, wherein the host cell is a microbial host cell.
21. A method for making an antibody-like binding protein comprising expressing in a cell one or more nucleic acid molecules encoding the antibody-like binding protein of any one of claims 1-7 or 9-17.
Dated this 30 day of April 2019
--Digitally Signed--
(Shakira N)
REG.No.IN/PA-972
Of De Penning & De Penning
Agent for the Applicant
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