Antigen binding polypeptides
The invention relates to a platform technology for production of antigen binding polypeptides having specificity for a desired target antigen which is based on the conventional antibody repertoire of species in the family Camelidae, and to antigen binding polypeptides obtained using this technology platform. In particular, the invention provides an antigen binding polypeptide comprising a VH domain and a VL domain, wherein at least one hypervariable loop or complementarity determining region (CDR) in the VH domain or the VL domain is obtained from a VH or VL domain of a species in the family Camelidae.
1. A method of modulating the binding of a human target antigen to its ligand or receptor comprising administration of an effective amount of a chimeric antigen binding polypeptide that specifically binds to a human target antigen to a subject, said polypeptide comprising a VH domain and a VL domain, said VH domain comprising hypervariable loops H1, H2 and H3, wherein said VH domain polypeptide is paired with a VL domain comprising hypervariable loops L1, L2and L3, wherein said VH domain and said VL domain are each fused to one or more IgG constant domains of a human antibody, wherein each of the hypervariable loops H1-H3 and L1-L3 are obtained from a conventional antibody of a Lama species by active immunization of the Lama species with the human target antigen, wherein said conventional antibody is a heterotetrameric IgG antibody composed of two identical light chains and two identical heavy chains, and wherein at least one of the hypervariable loops H1, H2, L1, L2 and L3 exhibits a predicted or actual canonical fold structure which is identical or substantially identical to a canonical fold structure of a corresponding H1, H2, L1, L2 or L3 hypervariable loop which occurs in a human antibody.
2. The method of claim 1 , wherein the human target antigen is one or more of: a receptor, receptor ligand, cell-signaling molecule, hormone, cytokine, cytokine receptor, or neurotransmitter.
3. The method of claim 2 , wherein the subject is a human.
4. The method of claim 1 , wherein the chimeric antigen binding polypeptide is formulated in a pharmaceutical dosage form.
5. The method of claim 1 , wherein the chimeric antigen binding polypeptide is formulated together with one or more pharmaceutically acceptable diluents, carriers, or excipients.
6. The method of claim 1 , wherein the chimeric antigen binding polypeptide is administered through one or more of the following routes: intravenous, intramuscular, intraperitoneal, intra-cerebro spinal, subcutaneous, intra-synovial, intrathecal, topical, sublingual, or inhalation.
7. The method of claim 1 , wherein the chimeric antigen binding polypeptide contains at least one amino acid substitution in at least one framework region of either the VH domain or the VL domain, in comparison to the VH and VL domain of a conventional antibody of a Lama species obtained by active immunization of the Lama species with the human target antigen.
8. The method of claim 1 , wherein the VH domain contains at least one amino acid sequence mismatch across the framework regions FR1, FR2, FR3 and FR4 in comparison with VH domains encoded by human germline or somatically mutated VH genes.
9. The method of claim 1 , wherein the VL domain contains at least one amino acid sequence mismatch across the framework regions FR1, FR2, FR3 and FR4 in comparison with VL domains encoded by human germline or somatically mutated VL genes.
10. The method of claim 1 , wherein the VH domain exhibits a sequence identity of 80% or greater with one or more human VH domains across the framework regions FR1, FR2, FR3 and FR4.
11. The method of claim 1 , wherein the VH domain exhibits a sequence identity of 90% or greater with one or more human VH domains across the framework regions FR1, FR2, FR3 and FR4.
12. The method of claim 1 , wherein the VH domain exhibits a sequence identity of 95% or greater with one or more human VH domains across the framework regions FR1, FR2, FR3 and FR4.
13. The method of claim 1 , wherein the VH domain exhibits a sequence identity of 97% or greater with one or more human VH domains across the framework regions FR1, FR2, FR3 and FR4.
14. The method of claim 1 , wherein the VL domain exhibits a sequence identity of 80% or greater with one or more human VL domains across the framework regions FR1, FR2, FR3 and FR4.
15. The method of claim 1 , wherein the VL domain exhibits a sequence identity of 90% or greater with one or more human VL domains across the framework regions FR1, FR2, FR3 and FR4.
16. The method of claim 1 , wherein the VL domain exhibits a sequence identity of 95% or greater with one or more human VL domains across the framework regions FR1, FR2, FR3 and FR4.
17. The method of claim 1 , wherein the VL domain exhibits a sequence identity of 97% or greater with one or more human VL domains across the framework regions FR1, FR2, FR3 and FR4.
18. The method of claim 1 , wherein hypervariable loop H1 and hypervariable loop H2in the VH domain form a canonical fold combination selected from the group consisting of:
1-1, 1-2, 1-3, 1-4, 1-6, 2-1, 3-1 and 3-5.
19. The method of claim 1 , wherein hypervariable loop L1 and hypervariable loop L2 in the VL domain form one of the following canonical fold combinations: 11-7, 13-7(A,B,C), 14-7(A,B), 12-11, 14-11, 12-12 , 2 -1, 3-1, 4-1 and 6-1.
20. The method of claim 1 , wherein the one or more IgG constant domains of a human antibody are of an isotype selected from the group consisting of: IgG, IgM, IgD, IgE, IgA.