Food Science and Technology Institute, Kyushu University 46-09, Hakozaki 6-10-1, 812, Higashi-ku, Fukuoka, Japan.
Cytotechnology. 1988 Jul;1(3):233-41. doi: 10.1007/BF00145026.
Human plasma was found to contain a macromolecular protein which can grow even a single cell of human lymphocytic cell lines (B-lymphoblastoid cell line HO-323-3 and T-lymphoblastoid cell line CCRF-CEM) and human-human hybridoma clones (SH-9, SU-1 and HB4C5) in a dish, but it has no effect on the growth of epithelial cell lines (lung cancer cell lines PC-8, QG-56 and QG-90). The proliferating activity for lymphocytic cell lines was gradually decreased at 4 or -20°C and dramatically decreased by heating at more than 60°C for 15 min. From human plasma, active fractions were purified by a successive application of Ca(2+) treatment, ammonium sulfate fractionation, DEAE-5PW column chromatography (FPLC) at pH 7.6. These active fractions were divided into at least three proteins by DEAE-5PW chromatography at pH 8.5 and chromatofocusing. These purified factors, named lymphocytic clonal growth factors (LCGFs), had similar molecular weights of about 600 K and each factor consisted of a 180 K and two 210 K subunits associated with hydrogen bondings. By the addition of 5 μg/ml of each factor into culture media, incidences of human-human hybridomas and cloning efficiencies of the hybridomas increased several-fold.
人血浆中存在一种高分子量蛋白，它能促进人淋巴细胞系（B-淋巴母细胞系 HO-323-3 和 T-淋巴母细胞系 CCRF-CEM）和人-人杂交瘤克隆（SH-9、SU-1 和 HB4C5）在培养皿中单个细胞的生长，但对上皮细胞系（肺癌细胞系 PC-8、QG-56 和 QG-90）的生长没有影响。淋巴细胞系的增殖活性在 4°C 或-20°C 时逐渐降低，在 60°C 以上加热 15 分钟时显著降低。从人血浆中，通过连续应用 Ca(2+)处理、硫酸铵分级、pH7.6 的 DEAE-5PW 柱层析（FPLC），可以纯化具有活性的组分。这些活性组分在 pH8.5 的 DEAE-5PW 层析和等电聚焦中被分为至少三种蛋白质。这些经纯化的因子被命名为淋巴细胞克隆生长因子（LCGFs），它们具有约 600 K 的相似分子量，每个因子由一个 180 K 和两个 210 K 亚基组成，这些亚基通过氢键相互连接。向培养基中加入 5μg/ml 的每种因子，人-人杂交瘤的发生率和杂交瘤的克隆效率增加了数倍。
