Assays for detecting neutralizing autoantibodies to biologic therapy

摘要

The present invention provides assays for detecting and measuring the presence or level of neutralizing and non-neutralizing autoantibodies to biologics such as anti-TNFα 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.

主权项

1. A method for measuring a percent of a neutralizing form of an autoantibody to an anti-TNFα drug in a sample, the method comprising:
(a) contacting the sample with a labeled anti-TNFα drug and a labeled TNFα to form:
(i) a first labeled complex of the labeled anti-TNFα drug and the autoantibody; and/or(ii) a second labeled complex of the labeled anti-TNFα drug, the labeled TNFα, 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 TNFα, free labeled anti-TNFα drug, and/or a complex of labeled anti-TNFα drug and labeled TNFα; (c) measuring an area under the curve (AUC) of free labeled TNFα in the sample (Free labeled TNFαAUC 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 TNFα measured in step (c) to an AUC of free labeled TNFα in a first control sample (Free labeled TNFαAUC of the first control sample) and an AUC of free labeled TNFα in a second control sample (Free labeled TNFαAUC of the second control sample) using the equation:
((Free labeled TNFαAUC of the sample−Free labeled TNFαAUC of the first control sample)/Free labeled TNFαAUC of the second control sample)*100,wherein the first control sample is a reference sample that is normal human serum to which labeled TNFα and labeled anti-TNFα drug has been added, and the second control sample is a reference sample that is normal human serum to which only labeled TNFα has been added.