A. Lischke et al., A CHEMILUMINESCENCE-BASED METHOD FOR THE DETECTION AND QUANTIFICATIONOF ANTIGEN-ANTIBODY INTERACTIONS ON THE SURFACE OF EUKARYOTIC CELLS, Analytical biochemistry, 236(2), 1996, pp. 322-326
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.