Quantitation of antibody binding to cell surface antigens by x-ray fluorescence spectrometry.
An X-ray spectrometric method has been developed to quantitate antibody bindng to whole cell surfaces in order to obtain a distribution of binding within a population of cells. The method involves incubation of target cells with ferritin-labeled antibody. Analysis of prepared samples in a modified transmission electron microscope with an X-ray detector and data analysis equipment, yields quantitative results on the binding of labeled antibody to individual cells. The binding of anti-2,4-dinitrophenol serum to Chinese hamster ovary cells with attached 2,4-dinitrophenol haptens was measured by X-ray spectrometry. Measurements of attached hapten by a radioisotopic marker correlated with the X-ray spectrometric determination of bound antibody. The use of synchronized cells in metaphase and G1 phases of the cell cycle permitted investigations into the binding per unit surface area. The distribution of antibody binding among a given population of cells was related to the surface area of the cells.
D000818:Animals; D000906:Antibodies; D000954:Antigens, Surface; D001666:Binding Sites, Antibody; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D006224:Cricetinae; D005260:Female; D010053:Ovary; D013052:Spectrometry, X-Ray Emission / Q000295:instrumentation / Q000379:methods
D000906:Antibodies; D000954:Antigens, Surface