Epitope binning investigates the sequential binding of two antibodies to an antigen. Using the BLI platform, antibodies are tested in pairs to determine the ability of an antibody to block another’s binding to the antigen. A blocking profile is created for each antibody against all other antibodies in the library. Antibodies with similar blocking profiles are grouped into a bin, indicating these antibodies bind to the same or overlapping epitopes.
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Antigen is immobilized onto a biolayer interferometry (BLI) sensor using an appropriate method. Antibody 1 is allowed to saturate its epitope on the antigen.
Binding of Antibody 2 to antigen saturated with Antibody 1 is assessed.
- 100 ug each of purified antibody
- 100 ug of purified antigen