Partial purification of murine B cell stimulatory factor (BSF)-1.
Partial purification of murine B cell stimulatory factor (BSF)-1.
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鼠 B 细胞刺激因子 (BSF)-1 的部分纯化。
DOI：
10.4049/jimmunol.135.4.2518
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发表时间：
1985
影响因子：
4.4
通讯作者：
W. Paul
中科院分区：
文献类型：
--
作者：
J. Ohara;S. Lahet;J. Inman;W. Paul
BSF-1 是从 EL-4 胸腺瘤系细胞的无血清培养物上清液中部分纯化的，该细胞系已在 48 小时前用 4 beta-phorbol-12 beta-myristate-12 alpha-acetate (PMA) 诱导。将 10 升批次的 BSF-1 吸附到三甲基硅基控孔玻璃珠 (TMS-CpG) 上并从其上洗脱，然后进行反相高效液相色谱 (RP-HPLC)。 TMS-CpG 和 RP-HPLC 的 BSF-1 活性回收率分别为 52% 至 55% 和 187% 至 227%。部分纯化的 BSF-1 的每毫克蛋白质的比活性比培养物上清液蛋白质的比活性高约 2600 倍。部分纯化的 BSF-1 具有 6.3 的单一等电点和表观分子量。通过等电聚焦和凝胶过滤-HPLC 分析时，分别在 18,000 和 21,700 之间。通过快速有效的程序制备大量部分纯化的 BSF-1 的能力应该对该淋巴因子的生化和免疫学研究有很大帮助。
BSF-1 was partially purified from serum-free culture supernatants of cells of the EL-4 thymoma line, which had been induced 48 hr earlier with 4 beta-phorbol-12 beta-myristate-12 alpha-acetate (PMA). BSF-1 in 10-liter batches was adsorbed onto and eluted from trimethylsilyl-controlled pore glass beads (TMS-CpG) and then subjected to reverse-phase high-performance liquid chromatography (RP-HPLC). The recovery of BSF-1 activity by TMS-CpG and RP-HPLC ranged from 52 to 55% and 187 to 227%, respectively. The specific activity in units per milligram of protein of partially purified BSF-1 was approximately 2600 times higher than that of the culture supernatant protein. The partially purified BSF-1 had a single isoelectric point of 6.3 and an apparent m.w. between 18,000 and 21,700 when analyzed by isoelectric focusing and gel filtration-HPLC, respectively. The ability to prepare large amounts of partially purified BSF-1 by a rapid and efficient procedure should be of great help in both biochemical and immunologic studies of this lymphokine.