Claims for Patent: 9,465,027

Summary for Patent: 9,465,027

Title:	Assays for detecting neutralizing autoantibodies to biologic therapy
Abstract:	The present invention provides assays for detecting and measuring the presence or level of neutralizing and non-neutralizing autoantibodies to biologics such as anti-TNF.alpha. drug therapeutics in a sample. The present invention is useful for monitoring the formation of neutralizing and/or non-neutralizing anti-drug antibodies over time while a subject is on biologic therapy. The present invention is also useful for predicting and/or determining the cross-reactivity of neutralizing anti-drug antibodies in a subject\'s sample with alternative biologic therapies. As such, the present invention provides information for guiding treatment decisions for those subjects receiving therapy with a biologic agent and improves the accuracy of optimizing therapy, reducing toxicity, and/or monitoring the efficacy of therapeutic treatment to biologic therapy.
Inventor(s):	Hauenstein; Scott (San Diego, CA), Ohrmund; Linda (San Diego, CA), Singh; Sharat (Rancho Santa Fe, CA), Wang; Shui Long (San Diego, CA)
Assignee:	Nestec S.A. (Vevey, CH)
Application Number:	14/144,261
Patent Claims:	1. A method for measuring a percent of a neutralizing form of an autoantibody to an anti-TNF.alpha. drug in a sample, the method comprising: (a) contacting the sample with a labeled anti-TNF.alpha. drug and a labeled TNF.alpha. to form: (i) a first labeled complex of the labeled anti-TNF.alpha. drug and the autoantibody; and/or (ii) a second labeled complex of the labeled anti-TNF.alpha. drug, the labeled TNF.alpha., and the autoantibody; (b) subjecting the first labeled complex and/or the second labeled complex to size exclusion chromatography to separate them from free labeled TNF.alpha., free labeled anti-TNF.alpha. drug, and/or a complex of labeled anti-TNF.alpha. drug and labeled TNF.alpha.; (c) measuring an area under the curve (AUC) of free labeled TNF.alpha. in the sample (Free labeled TNF.alpha..sub.AUC of the sample) after size exclusion chromatography; and (d) measuring the percent of a neutralizing form of the autoantibody by comparing the AUC of free labeled TNF.alpha. measured in step (c) to an AUC of free labeled TNF.alpha. in a first control sample (Free labeled TNF.alpha..sub.AUC of the first control sample) and an AUC of free labeled TNF.alpha. in a second control sample (Free labeled TNF.alpha..sub.AUC of the second control sample) using the equation: ((Free labeled TNF.alpha..sub.AUC of the sample-Free labeled TNF.alpha..sub.AUC of the first control sample)/Free labeled TNF.alpha..sub.AUC of the second control sample)*100, wherein the first control sample is a reference sample that is normal human serum to which labeled TNF.alpha. and labeled anti-TNF.alpha. drug has been added, and the second control sample is a reference sample that is normal human serum to which only labeled TNF.alpha. has been added. 2. The method of claim 1, wherein the anti-TNF.alpha. drug is selected from the group consisting of infliximab, etanercept, adalimumab, certolizumab pegol, golimumab, and combinations thereof. 3. The method of claim 1, wherein the autoantibody to the anti-TNF.alpha. drug is selected from the group consisting of a human anti-chimeric antibody (HACA), a human anti-humanized antibody (HAHA), a human anti-mouse antibody (HAMA), and combinations thereof. 4. The method of claim 1, wherein the sample is positive for the neutralizing form of the autoantibody when the sample has greater than or equal to about 3.00% of the neutralizing form of the autoantibody. 5. The method of claim 1, wherein the sample is serum. 6. The method of claim 1, wherein the sample is obtained from a subject receiving therapy with the anti-TNF.alpha. drug. 7. The method of claim 1, wherein the labeled anti-TNF.alpha. drug and the labeled TNF.alpha. comprise different fluorophores or fluorescent dyes. 8. A method for determining whether a neutralizing form of an autoantibody to a first anti-TNF.alpha. drug is cross-reactive with a second anti-TNF.alpha. drug, the method comprising: (a) measuring the percent of a neutralizing form of the autoantibody in a sample by performing the method of claim 4 to determine whether the sample is positive or negative for the neutralizing form of the autoantibody; and if the sample is positive for the neutralizing form of the autoantibody, then: (b) contacting the sample with a labeled second anti-TNF.alpha. drug to form a labeled complex of the labeled second anti-TNF.alpha. drug and the neutralizing form of the autoantibody; (c) subjecting the labeled complex to size exclusion chromatography to separate the labeled complex; and (d) detecting the labeled complex, thereby determining whether a neutralizing form of an autoantibody to a first anti-TNF.alpha. drug is cross-reactive with a second anti-TNF.alpha. drug. 9. The method of claim 8, wherein the first and second anti-TNF.alpha. drugs are independently selected from the group consisting of infliximab, etanercept, adalimumab, certolizumab pegol, golimumab, and combinations thereof. 10. The method of claim 8, wherein the autoantibody to the first anti-TNF.alpha. drug is selected from the group consisting of a human anti-chimeric antibody (HACA), a human anti-humanized antibody (HAHA), a human anti-mouse antibody (HAMA), and combinations thereof. 11. The method of claim 8, wherein the presence of the labeled complex indicates that the neutralizing autoantibody against the first anti-TNF.alpha. drug is cross-reactive with the second anti-TNF.alpha. drug. 12. The method of claim 8, wherein the absence of the labeled complex indicates that the neutralizing autoantibody against the first anti-TNF.alpha. drug is not cross-reactive with the second anti-TNF.alpha. drug. 13. The method of claim 8, wherein the sample is serum. 14. The method of claim 8, wherein the sample is obtained from a subject receiving therapy with the anti-TNF.alpha. drug. 15. The method of claim 8, wherein the labeled second anti-TNF.alpha. drug comprises a fluorophore or fluorescent dye.

Details for Patent 9,465,027

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Janssen Biotech, Inc.	REMICADE	infliximab	For Injection	103772	08/24/1998		2031-07-06
Immunex Corporation	ENBREL	etanercept	For Injection	103795	11/02/1998		2031-07-06
Immunex Corporation	ENBREL	etanercept	For Injection	103795	05/27/1999		2031-07-06
Immunex Corporation	ENBREL	etanercept	Injection	103795	09/27/2004		2031-07-06
Immunex Corporation	ENBREL	etanercept	Injection	103795	02/01/2007		2031-07-06