A CHEMILUMINESCENCE-BASED METHOD FOR THE DETECTION AND QUANTIFICATIONOF ANTIGEN-ANTIBODY INTERACTIONS ON THE SURFACE OF EUKARYOTIC CELLS

Enhanced chemiluminescence was applied to detect the binding of monocl onal antibodies to surface antigens on intact cells. The fast and simp le assay is performed in the microtiter scale and thus allows for the simultaneous processing of a large number of samples with a sensitivit y comparable to conventionally used techniques such as cytometry or We stern blot analysis. In two model experiments, we demonstrate (a) the detection of a heterologously expressed cytokine receptor subunit on t he surface of suspension cells and (b) the screening of hybridoma clon es for the production of antibodies specifically recognizing surface a ntigens on a tumor cell line. Moreover, the assay is shown to be suita ble for the determination of antibody affinities and of antibody bindi ng sites per cell. (C) 1996 Academic Press, Inc.