Claims for Patent: 10,261,094

Summary for Patent: 10,261,094

Title:	Competitive ligand binding assay for detecting neutralizing antibodies
Abstract:	A immunogenicity assay for detecting the presence of neutralizing antibodies to a biotherapeutic protein wherein the biotherapeutic protein has been administered to a patient in need, comprising the steps of (a) obtaining a sample from the patient; (b) incubating the sample in the presence of a capture reagent; and (c) adding a detecting reagent, wherein a decreased signal relative to a control sample indicates the presence of a neutralizing antibody to the biotherapeutic agent.
Inventor(s):	Rajadhyaksha; Manoj (Colchester, CT), Partridge; Michael (Eastchester, NY), Torri; Albert (Lagrangeville, NY)
Assignee:	Regeneron Pharmaceuticals, Inc. (Tarrytown, NY)
Application Number:	15/031,126
Patent Claims:	1. A method for detecting the presence of neutralizing antibodies to a biotherapeutic protein, wherein the biotherapeutic protein has been administered to a patient in need, comprising the steps of (a) decreasing the pH of the patient sample to an acidic pH, (b) combining the patient sample with an antibody that binds to a target of the biotherapeutic protein and a capture reagent comprising a labeled biotherapeutic protein, wherein the label on the biotherapeutic protein facilitating binding of the biotherapeutic protein to a surface, and (c) adding a detecting reagent comprising a labeled target of the biotherapeutic protein, wherein a decreased signal relative to a control sample indicates the presence of a neutralizing antibody to the biotherapeutic protein. 2. The method of claim 1, wherein the protein biotherapeutic is a monoclonal antibody. 3. The method of claim 2, wherein the protein biotherapeutic is an anti-interleukin-6 receptor .alpha. (IL-6R.alpha.) monoclonal antibody. 4. The method of claim 3, wherein the protein biotherapeutic is sarilumab or tocilizumab. 5. The method of claim 4, wherein the protein biotherapeutic is sarilumab. 6. The method of claim 1, wherein the patient is treated for an IL-6-dependent disease. 7. The method of claim 6, wherein the IL-6-dependent disease is one of rheumatoid arthritis, diabetes, atherosclerosis, Alzheimer's disease, systemic lupus erythematosus, multiple myeloma, connective tissue disorders, Castleman's disease and prostate cancer. 8. The method of claim 7, wherein the IL-6-dependent disease is rheumatoid arthritis. 9. The method of claim 1, wherein the capture reagent is labeled sarilumab. 10. The method of claim 9, wherein the labeled sarilumab is biotinylated sarilumab. 11. The method of claim 10, wherein the biotinylated sarilumab is bound to an avidin-coated plate. 12. The method of claim 1, wherein the target is soluble IL-6 receptor .alpha.. 13. The method of claim 1, wherein the labeled target is ruthenium labeled soluble IL-6 receptor .alpha.. 14. The method of claim 1, wherein the pH of the patient sample is decreased to an acidic pH by treatment with an acidic solution. 15. The method of claim 1, further comprising increasing the pH of the patient sample to a neutral pH following the decrease of the patient sample to an acidic pH. 16. The method of claim 1, wherein the method exhibits a sensitivity tolerance of about 150 ng/mL, a drug tolerance of about 500 ng/mL, and a target interference tolerance of about 1 .mu.g/mL. 17. A method for detecting the presence of neutralizing antibodies to sarilumab, wherein sarilumab has been administered to a patient in need, comprising the steps of (a) decreasing the pH of a serum sample to an acidic pH; (b) combining a patient serum sample with a capture reagent comprising biotinylated sarilumab; (c) increasing the pH of the combined serum sample and capture reagent to a neutral pH; and (d) adding a detecting reagent, wherein the detecting reagent comprises labeled soluble IL-6 receptor .alpha., wherein a decreased signal relative to a control sample indicates the presence of a neutralizing antibody to sarilumab. 18. The method of claim 17, wherein the detecting reagent is ruthenium labeled soluble IL-6 receptor .alpha.. 19. The method of claim 17, wherein the patient is suffering from rheumatoid arthritis. 20. The method of claim 17, wherein the method exhibits a sensitivity tolerance of about 150 ng/mL, a drug tolerance of about 500 ng/mL, and a target interference tolerance of about 1 .mu.g/mL. 21. The method of claim 14, wherein the acidic solution is acetic acid. 22. The method of claim 15, wherein the pH is increased to a neutral pH by treatment with a basic solution. 23. The method of claim 22, wherein the basic solution is a Tris base. 24. The method of claim 17, wherein the pH is decreased to an acidic pH by treatment with an acid solution. 25. The method of claim 24, wherein the acidic solution is acetic acid. 26. The method of claim 17, wherein the pH is increased to a neutral pH by treatment with a basic solution. 27. The method of claim 26, wherein the basic solution is a Tris base. 28. The method of claim 1, wherein the label of the labeled biotherapeutic-protein is biotin. 29. The method of claim 1, wherein the antibody binds free target of the biotherapeutic protein released by the low pH treatment and mitigates target interference.

Details for Patent 10,261,094

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	ACTEMRA	tocilizumab	Injection	125276	01/08/2010		2033-10-31
Genentech, Inc.	ACTEMRA	tocilizumab	Injection	125472	10/21/2013		2033-10-31
Genentech, Inc.	ACTEMRA	tocilizumab	Injection	125472	11/19/2018		2033-10-31
Sanofi-aventis U.s. Llc	KEVZARA	sarilumab	Injection	761037	05/22/2017		2033-10-31
Sanofi-aventis U.s. Llc	KEVZARA	sarilumab	Injection	761037	04/13/2018		2033-10-31