Purification of functional subunits of the eighth component of human complement (C8) under nondenaturing conditions.
Purification of functional subunits of the eighth component of human complement (C8) under nondenaturing conditions.
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在非变性条件下纯化人类补体第八种成分 (C8) 的功能亚基。
DOI：
10.1159/000467835
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
Sodetz,JM
中科院分区：
文献类型：
--
作者：
Rao,AG;Sodetz,JM
本文描述了一种简单的方法，在不加变性剂的条件下，以明显高于以前的产率纯化人C8的非共价结合的α-γ和β亚基。该方法在1.5 M NaCl存在下采用常规或高压分子筛色谱法。这种离子强度促进α-γ与β的解离，并促进它们在温和条件下的物理分离。产率通常为60-75%，并且根据几个标准判断，每个亚基在功能上与通过使用十二烷基硫酸钠的早期方法分离的那些亚基相同。这种新方法消除了对这种洗涤剂的需要，从而避免了在其去除过程中遇到的主要溶解度和产率问题。
A simple procedure is described for purifying the noncovalently associated α-γand β subunits of human C8 in the absence of dénaturants and in yields significantly higher than previously obtainable. The procedure utilizes either conventional or high-pressure molecular sieve chromatography in the presence of 1.5 M NaCl. This ionic strength promotes dissociation of α-γ from β and facilitates their physical separation under mild conditions. Yields are typically 60-75% and, as judged by several criteria, each subunit is functionally identical to those isolated by earlier methods using sodium dodecyl sulfate. This new procedure eliminates the need for this detergent and thereby avoids the major solubility and yield problems encountered during its removal.