Partial purification of human lymphocyte activating factor.
Partial purification of human lymphocyte activating factor.
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人淋巴细胞激活因子的部分纯化。
DOI：
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发表时间：
1980
期刊：
影响因子：
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通讯作者：
R. Metzgar
中科院分区：
文献类型：
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作者：
L. Lachman;S. Page;R. Metzgar
淋巴细胞激活因子(LAF)是人单核细胞在含有5%人血清和非特异性免疫刺激剂脂多糖的组织培养液中培养18-24小时后释放的一种T淋巴细胞刺激剂。LAF的纯化实质上是从生产活性所需的人血清蛋白中分离出低分子量的LAF(约13,000)。中空纤维超滤法可以快速分离血清蛋白中的低分子量LAF，但产率仅为原活性的20%。等电聚焦(IEF)有效地将LAF从所有微量的人血清中分离出来，导致纯化的样品不包含可测量的蛋白质，也不显示聚丙烯酰胺凝胶上的条带。纯化后的IEF活性约为纯培养基中低相对分子质量活性的2%，在生物检测系统中具有很高的活性。
Lymphocyte Activating Factor (LAF) is a T lymphocyte stimulant released by human monocytes cultured for 18-24 hours in tissue culture medium containing 5% human serum and the non-specific immunostimulant lipopolysaccharide. The purification of LAF is essentially the separation of low MW LAF (approximately 13,000) from the human serum proteins required for production of the activity. Hollow fiber ultrafiltration has been found to effect a rapid separation of low MW LAF from serum proteins, but with a yield of only 20% of the original activity. Isoelectric focusing (IEF) efficiently separates LAF from all traces of human serum, resulting in a purified sample containing no measurable protein and revealing no bands on polyacrylamide gels. The IEF purified material is about 2% of the low MW activity present in the unfractionated culture medium and is highly active in the biological assay system.