Binding of cells to matrixes of distinct antibodies coated on solid surface

The present studies investigate the potential of simultaneous multiple determinations of specific cell surface antigens in one reaction incubation by employing orderly arranged antibody spots on a solid surface. Antibodies of distinct specificities were coated on very small areas in close proximity forming matrix-like arrays on glass cover slips. These antibody spots were found to be capable of serving as minute specific immunoadsorbents for cells bearing on their surface the antigens with which the antibodies reacted. Antibody spots of 1.0, 0.5 and 0.25 mm diameter could adsorb maximally about 17,000, 4500, and 1100 mononuclear cells. An area of 1 cm2 could be coated with 25, 100, or 400 of these spots, respectively. In matrixes that contained anti-Lyt 2.1 and anti-Lyt 2.2 antibody spots, AKR (Lyt 2.1+) thymocytes adhered only to anti-Lyt 2.1 spots, BALB/c (Lyt 2.2+) thymocytes only to anti-Lyt 2.2 spots, and thymocytes of (AKR X BALB/c) F1 to both spots. The potential of this method for determining allotypes of HLA antigens and for determining in a mixed cell population the proportions of subsets bearing specific differentiation antigens is discussed.