Antigen-Specific Cell Adherence Assay: A New Method for Separation of Antigen-Specific Hybridoma Cells

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Abstract
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.