Epitope binning of murine monoclonal antibodies by a multiplexed pairing assay

We describe an assay and data evaluation technique for sorting a panel of murine monoclonal antibodies according to epitope specificities. The assay analyzes the simultaneous binding (pairing) of antibodies to an antigen and groups together antibodies with similar pairing profiles. Similar profiles indicate that the antibodies bind to the same or closely related epitopes. The assay works well with crude hybridoma supernatants and can be multiplexed. These features make the assay particularly suitable for the early phase of hybridoma/antibody screening when antibodies are available only as low volume culture harvest samples.