Partial purification of human lymphocyte-activating factor (LAF) by ultrafiltration and electrophoretic techniques.
Partial purification of human lymphocyte-activating factor (LAF) by ultrafiltration and electrophoretic techniques.
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通过超滤和电泳技术部分纯化人淋巴细胞激活因子（LAF）。
DOI：
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发表时间：
1977
影响因子：
4.4
通讯作者：
R. Handschumacher
中科院分区：
文献类型：
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作者：
L. Lachman;M. Hacker;R. Handschumacher
淋巴细胞激活因子（LAF）是通过在脂多糖和自体人血清存在下培养人外周血白细胞而产生的。主要的 LAF 活性可通过 Sephadex 色谱法（分子量 13,000）通过中空纤维装置超滤与培养基中的大多数血清蛋白分离。浓缩超滤液的蔗糖梯度等电聚焦产生 LAF 活性的单峰，平均等电点为 pH 6.8。等电聚焦样品的半制备型聚丙烯酰胺凝胶电泳导致 LAF 活性与可检测量的血清蛋白分离。回收的 LAF 活性估计纯化超过 16,000 倍，并被判断为在亚微克量下具有活性。
Lymphocyte-activating factor (LAF) has been produced by culturing human peripheral blood leukocytes in the presence of lipopolysaccharide and autologous human serum. The major LAF activity, identifiable by Sephadex chromatography (m.w. 13,000) was separated from most serum proteins in the culture medium by ultrafiltration with a hollow fiber device. Sucrose gradient isoelectric focusing of the concentrated ultrafiltrate yielded a single peak of LAF activity with an average isoelectric point of pH 6.8. Semi-preparative polyacrylamide gel electrophoresis of the isoelectric focusing sample resulted in separation of the LAF activity from detectable amounts of serum proteins. The recovered LAF activity was estimated to be purified more than 16,000-fold and is judged to be active in submicrogram amounts.