title: Human monoclonals from antigen-specific selection of B lymphocytes and transformation by EBV.
abstract: Hybridoma technology has made it possible to prepare monoclonal antibodies with the use of murine lymphocytes. Attempts to extend this technology to the human level, however, have met with difficulties. A method has been developed for making human monoclonal antibodies of predetermined specificity. Biotinylated antigens (human thyroglobulin or tetanus toxoid) were incubated with human B lymphocytes from peripheral blood. The lymphocytes to which the antigens bound were selected by fluorescence-activated cell sorting. Positively selected (high fluorescence) and negatively selected (low fluorescence) cells were then transformed with Epstein-Barr virus (EBV) and grown in microculture wells. All wells from the positively selected fraction produced antigen-specific antibody (95 to 1800 nanogram-equivalents per milliliter), whereas fewer than 6% of the wells from negatively selected fraction made any detectable antibody (less than 10 nanogram-equivalents per milliliter). When the positively selected EBV-transformed cells were cultured in limiting dilution, clones were obtained that made antigen-specific monoclonal antibodies. By this method, monoclonal antibodies to both foreign antigens and autoantigens can be prepared from the normal human B-cell repertoire.
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002469:Cell Separation / Q000379:methods; D002472:Cell Transformation, Viral; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology; D005434:Flow Cytometry; D004854:Herpesvirus 4, Human; D006801:Humans; D007075:Immunoglobulin M / Q000096:biosynthesis; D011947:Receptors, Antigen, B-Cell / Q000276:immunology; D013745:Tetanus Toxoid / Q000276:immunology; D013954:Thyroglobulin / Q000276:immunology; D013967:Thyroiditis, Autoimmune / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M; D011947:Receptors, Antigen, B-Cell; D013745:Tetanus Toxoid; D013954:Thyroglobulin