Panning separation for monoclonal antibody-specific T-cell subsets: effects of enzymes and metabolic inhibitors.
The mechanisms of "panning," a simple positive cell separation technique, were examined. Using monoclonal antibodies to "pan" for T cells (T101+, T4+, and T8+), we obtained enriched populations with 90% purity and viability from unfractionated human peripheral blood lymphocytes (PBL). We found that the "panning" method reflects an active process. It is sodium azide inhibitable and independent of the divalent cation concentration. Effective panning does not require capping, patch formation, or DNA synthesis. The cell yields are unaffected by mitomycin-C or microtubule and microfilament blocking reagents such as concanavalin A, colchicine, and cytochalasin B. This economical technique provides large numbers of functionally intact monoclonal antibody-specific cells within a relatively short time for further functional and biochemical characterization.
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001386:Azides / Q000494:pharmacology; D002469:Cell Separation / Q000379:methods*; D003078:Colchicine / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D005260:Female; D006801:Humans; D007163:Immunosorbent Techniques; D008297:Male; D051379:Mice; D011817:Rabbits; D019810:Sodium Azide; D013601:T-Lymphocytes / Q000145:classification* / Q000187:drug effects / Q000276:immunology
D000911:Antibodies, Monoclonal; D001386:Azides; D003571:Cytochalasin B; D019810:Sodium Azide; D003078:Colchicine