Removal of phytohemagglutinin from conditioned medium by affinity chromatography.
Supernatants of lymphocytes cultured with phytohemagglutinin (PHA-induced conditioned medium) are known to contain residual lectin. Studies with T cells specifically sensitized against a given antigen and maintained in culture by the T cell growth factor in conditioned medium may be hampered by the presence of PHA since the lectin could induce polyclonal activation of T cells. We developed a procedure for removing lectin from conditioned medium by affinity adsorption on porcine thyroglobulin-Sepharose. The affinity method was capable of removing detectable amounts of lectin since the mitogenic capacity for peripheral blood lymphocytes was lost after adsorption. In contrast, thyroglobulin-Sepharose adsorbed CM retained good mitogenic activity and growth-supporting capacity for human cultured T cells.
D000327:Adsorption; D000818:Animals; D002455:Cell Division; D002846:Chromatography, Affinity; D003470:Culture Media; D006133:Growth Substances / Q000494:pharmacology; D006801:Humans; D007164:Immunosorbents; D010835:Phytohemagglutinins; D012685:Sepharose / Q000378:metabolism; D013552:Swine; D013601:T-Lymphocytes / Q000166:cytology; D013954:Thyroglobulin / Q000378:metabolism
D003470:Culture Media; D006133:Growth Substances; D007164:Immunosorbents; D010835:Phytohemagglutinins; D013954:Thyroglobulin; D012685:Sepharose