Antigen-specific cell adherence assay: a new method for separation of antigen-specific hybridoma cells.
A new method for the detection and separation of antigen-specific antibody-producing cells on the basis of antibody-mediated recognition of solid-phase immobilized antigen molecules is described. Hybridoma cells are placed on microtiter plate wells coated with antigen molecules, and antigen-specific antibody-producing cells bind to the immobilized antigen molecules; antibody nonproducing or nonspecific antibody-producing cells can be easily separated from the bound cells by inverting the plate. Cells bound to solid-phase immobilized antigen molecules can readily be quantitated by counting under a light microscope, and the cells recovered can produce antibody in culture. Unspecific binding of cells in antigen-specific cell adherence assay (ASCAA) is optimally below 5%. Also, effect of drugs interfering with processes related to antibody production of antigen-specific cells can be detected and evaluated by ASCAA.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D000921:Antibody-Producing Cells / Q000166:cytology* / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000032:analysis; D000941:Antigens / Q000276:immunology; D002448:Cell Adhesion; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000166:cytology*; D051379:Mice
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000941:Antigens