Antigen‐specific adherence of antibody‐producing hybridoma cells: Application to neural and nonneural antigens

Abstract

Antibody-producing hybridoma cells specifically bind to microgram quantities of antigen molecules adsorbed onto the surface of plastic microtiter plates. The binding of hybridoma cells to nonantigen is optimally below 5%, similar binding of non-antibody-producing cells is 4-7%, compared to the binding of the hybridomas to their antigen. There is a difference in the kinetics of binding hybridomas to antigen compared to nonantigen. The number of bound cells depends on the amount, i.e., the surface density, of the antigen molecules and shows typical saturation effects. Preincubation of hybridomas with excess free antigen and saturation of the antibody binding site on the surface with the hybridoma-produced antibody reduces binding of the hybridoma cells to the antigen. Treatment of cells with trypsin reduces binding to antigen-coated plastic surfaces. Drugs such as sodium azide, cytochalasin B, colchicine, vinpocetine, and vincristine sulfate reduce binding to the antigen. Hybridoma cells adhering to the antigen produce more antibody than nonadhering cells. The results reported in this paper show that antigen molecules adsorbed to include a plastic surface and hybridoma cells interact specifically. This system forms a suitable model to study the interaction of antigen with antigen-specific cells and may be useful as a separation method for specific antibody-producing cells.

B-T lymphocyte interactions in experimental autoimmune myasthenia gravis: antigen presentation by ratlmouse hybridoma lines secreting monoclonal antibodies against the nicotinic acetylcholine receptor

“…Then, competition for soluble antigens between secreted immunglobulins antibodies and cell surface-bound antigen receptors could be expected to interfere with binding of the antigen and its subsequent incorporation. This is, however, not the case, as demonstrated by the formation of stable rosettes between antigen-presenting hybridomas and recognizing T cells in our experiments, and by adsorption of hybridomas to insolubilized antigen [30].…”