Detection of biomarkers and biomarker complexes

摘要

The invention features methods and devices for the detection of biomarker complexes and their components and for the sequential detection of multiple epitopes of a biomarker. The invention also features methods for diagnosing disease and evaluating the efficacy of treatment of a subject with a disease.

主权项

1. A method of sequentially detecting three epitopes of a biomarker in a biological sample, said method comprising:
(a) contacting said biological sample with a device having an immobilized binding agent on the surface thereof in a pattern that generates a signal so that said biomarker in said sample specifically binds to said immobilized binding agent, (b) contacting said surface with a first detecting binding agent, wherein said first detecting binding agent specifically binds to a first epitope of the biomarker and not to a second or third epitope of the biomarker; (c) detecting said first epitope by a change in signal produced by said pattern upon binding of said first detecting binding agent to said first epitope; (d) contacting said surface with a second detecting binding agent, wherein said second detecting binding agent specifically binds to the second epitope of said biomarker and does not specifically bind to said first or third epitope; (e) detecting said second epitope by a change in signal produced by said pattern upon binding of said second detecting binding agent to said second epitope; (f) contacting said surface with a third detecting binding agent, wherein said third detecting binding agent specifically binds to the third epitope of said biomarker and does not specifically bind to said first or second epitope; and (g) detecting said third epitope by a change in signal produced by said pattern upon binding of said third detecting binding agent to said third epitope.