Rapid generation of anti-idiotypic antibodies
View Patent ↗The present disclosure relates to in vivo methods for producing anti-idiotypic antibodies. In some aspects, anti-idiotypic antibodies are generated by co-administering to a mouse a first antibody having a murine IgG2a isotype and a second antibody that targets mouse B cells and has a murine IgG2a isotype. In some embodiments, the mouse expresses the Igh-1 b allele of IgG2a, and the second antibody binds a mouse B cell surface marker selected from the group consisting of CD19, CD20, CD21, CD22, CD40, CD45, IgM, and IgD.
1. A method for producing anti-idiotypic antibodies comprising:
(a) co-administering to a mouse a first antibody having a murine IgG2a isotype and a second antibody that targets mouse B cells and has a murine IgG2a isotype; wherein the first and second antibodies have different binding specificities, wherein the first antibody is soluble, wherein the mouse expresses the Igh-1 b allele of IgG2a, and wherein the second antibody binds a mouse B cell surface marker selected from the group consisting of CD19, CD20, CD21, CD22, CD40, CD45, IgM, and IgD;
(b) producing a hybridoma fusion of spleen cells from the immunized mouse and myeloma fusion partners; and
(c) isolating the anti-idiotypic antibodies which specifically bind to the first antibody in step (a).
2. The method of claim 1 , wherein said mouse is susceptible to autoimmune disease.
3. The method of claim 1 , wherein the myeloma fusion partners are either NS-1 or SP2/0 cells.
4. The method of claim 1 , wherein said mouse is selected from the group consisting of: non-obese diabetic (NOD), non-obese resistant (NOR), SJL, C.B-17, and C57BL/6.
5. The method of claim 4 , wherein said mouse is a NOD mouse.
6. The method of claim 1 , wherein said co-administration is performed sequentially.
7. The method of claim 6 , wherein the sequential co-administrations are given as boosting administrations.
8. The method of claim 1 , wherein said co-administration is performed simultaneously.
9. The method of claim 1 , wherein said first and second antibody are administered at a ratio of about 1:1.
10. The method of claim 1 , wherein said first and second antibody are administered at a ratio of about 1:2.
11. The method of claim 1 , wherein said first and second antibody are administered at a ratio of about 1:4.
12. The method of claim 1 , wherein said second antibody is the anti-mCD20 antibody 18B 12.
13. The method of claim 1 , wherein said first antibody specifically binds an antigen selected from the group consisting of: alpha-4-integrin, glycoprotein I1b/I11a, vascular endothelial growth factor, epidermal growth factor, complement C5 protein, ErbB2, CD3 receptor, CD11a, CD20, CD23, CD25, CD33, CD52, BCMA, CD40, Lymphotoxin α, Lymphotoxin α 1 β 2 , LIGHT, TWEAK, CD154, VLA4, EGFR, IGF1R, CD169, IL-6, IL-23, TNF-α, Neonatal Fe Receptor (FcRn), BDCA-2, DCIR, DR6 (Death Receptor 6), LINGO-I, Tyro3, RON receptor tyrosine kinase, DDR1 (Discoidin Domain Receptor 1), HER3, FN14, VEGF and CD103.
14. The method of claim 13 , wherein said first antibody comprises the variable domains of rituximab.
15. A method for producing anti-idiotypic antibodies comprising:
(a) co-administering to a mouse a first antibody having a murine lgG2a isotype and a second antibody that targets the mouse B cell surface marker CD20 and has a murine IgG2a isotype, wherein the first and second antibodies have different binding specificities, wherein the first antibody is soluble, wherein the mouse expresses the lgh-1 b allele of IgG2a;
(b) producing a hybridoma fusion of spleen cells from the immunized mouse and myeloma fusion partners; and
(c) isolating the anti-idiotypic antibodies which specifically bind to the first antibody in step (a).
16. The method of claim 15 , wherein said mouse is susceptible to autoimmune disease.
17. The method of claim 15 , wherein the myeloma fusion partners are either NS-1 or SP2/0 cells.
18. The method of claim 15 , wherein said mouse is selected from the group consisting of: non-obese diabetic (NOD), non-obese resistant (NOR), SJL, C.B-17, and C57BL/6.
19. The method of claim 15 , wherein said mouse is a NOD mouse.
20. The method of claim 15 , wherein said co-administration is performed sequentially.
21. The method of claim 20 , wherein the sequential co-administrations are given as boosting administrations.
22. The method of claim 15 , wherein said co-administration is performed simultaneously.
23. The method of claim 15 , wherein said first and second antibody are administered at a ratio of about 1:1 , about 1:2, or about 1:4.
24. The method of claim 15 , wherein said first antibody specifically binds an antigen selected from the group consisting of: alpha-4-integrin, glycoprotein I1b/I11a, vascular endothelial growth factor, epidermal growth factor, complement CS protein, ErbB2, CD3 receptor, CD11a, CD20, CD23, CD25, CD33, CD52, BCMA, CD40, Lymphotoxin α, Lymphotoxin α 1 β 2 , LIGHT, TWEAK, CD I 54, VLA4, EGFR, IGFI R, CD169, IL-6, IL-23, TNF-α, Neonatal Fe Receptor (FcRn), BDCA-2, DCIR, DR6 (Death Receptor 6), LINGO-I , Tyro3, RON receptor tyrosine kinase, DDR1 (Discoidin Domain Receptor 1), HER3, FN14, VEGF and CD103.
25. The method of claim 24 , wherein said first antibody comprises the variable domains of rituximab.