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Panning Separation for Monoclonal Antibody‐Specific T‐Cell Subsets: Effects of Enzymes and Metabolic Inhibitors
Author(s) -
Goto Makoto,
Bluestein Harry G.,
Zvaifler Nathan J.
Publication year - 1984
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1984.tb00795.x
Subject(s) - panning (audio) , monoclonal antibody , biology , cytochalasin b , microbiology and biotechnology , concanavalin a , sodium azide , antibody , microfilament , enzyme , cell , biochemistry , cytoskeleton , immunology , in vitro , paleontology , zoom , lens (geology)
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.