Purification of human B cell growth factor.
Human B cell growth factor (BCGF, 12,000 to 14,000 daltons) has been purified from lectin-stimulated, peripheral blood mononuclear cell-conditioned medium. The purification procedure involves a series of column chromatographic steps incorporating ion exchange, affinity binding, and gel filtration. This procedure is centered around a relatively high yield single chromatographic step, for the removal of co-eluting cytokines from BCGF, that is based on differential binding characteristics to the weak ion-exchange matrix, hydroxylapatite. Reverse-phase high-pressure liquid chromatographic separation on a C18-Bondapak column effectively separates the BCGF and TCGF moieties, yet is characterized by poor yields. High-pressure liquid chromatographic procedures on anion exchange and size exclusion provided the final purification step for BCGF, at an analytical level, resulting in a single band with a m.w. of 12,000 on a SDS-polyacrylamide gel.
D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D003470:Culture Media / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006133:Growth Substances / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D007376:Interleukin-2 / Q000302:isolation & purification / Q000378:metabolism; D015847:Interleukin-4; D008222:Lymphokines / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D009000:Monocytes / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate; D013601:T-Lymphocytes / Q000378:metabolism
D003470:Culture Media; D006133:Growth Substances; D007376:Interleukin-2; D008222:Lymphokines; D015847:Interleukin-4; D012967:Sodium Dodecyl Sulfate