人B细胞生长因子的分离提纯

Biochemical separation of human B cell growth factor

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 the present report a protocal for the purification of human B cell growth factor (12 KD-BCGF)has been described. Conditioned medium from lectin-stimulated T cells was fractioned by ammonium sulfate precipitation, ion-exchange chromatography, affinity binding chromatography and gel filtration chromatography. Our precedures take the advantage of the differential binding properties of BCGF and IL-2 to the weak ion-exchange matrix hydroxylapatite for an effective removal of IL-2 from BCGF. A BCGF-responsive B cell line 3D5 was used astarget cells for the assay of BCGF activity. SDS-PAGE analysis shows three protein bands in the Bio-Gel P-30 eluted materials, with a BCGF activity in the 12KD band.