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本工作用对B细胞生长因子(BCGF)有特异反应的3D5细胞检测BCGF活性,并利用对弱离子交换介质羟基磷灰石结合特性的不同,有效地从有丝分裂原刺激的人T细胞培养上清液中除去IL-2,得到了部分纯化的BCGF。最终凝胶过滤组份经SDS-PAGE分析显示有三条蛋白带,其中12KD蛋白带有BCGF活性。电泳分析比较表明,从制备低分子量BCGF这点来看,我们分离到的BCGF优于Cellular Product公司生产的cpBCGF(产品号16189)。
In this work, we detected the activity of BCGF by using 3D5 cells that have a specific response to B cell growth factor (BCGF). Based on the binding properties of hydroxyapatite, which is a weak ion exchange medium, we effectively isolated the mitogen-stimulated human T cell culture supernatant IL-2 was removed from the fluid to obtain partially purified BCGF. The final gel filtration fraction showed three protein bands by SDS-PAGE analysis, with 12KD protein carrying BCGF activity. Electrophoretic analysis shows that BCGF we isolated is superior to cpBCGF (product number 16189) produced by Cellular Product from the viewpoint of preparation of low molecular weight BCGF.