Purification of the first component of human complement by affinity chromatography on human globulin linked to sepharose.
Purification of the first component of human complement by affinity chromatography on human globulin linked to sepharose.
复制标题
通过对与琼脂糖凝胶连接的人球蛋白进行亲和层析来纯化人补体的第一成分。
DOI：
10.4049/jimmunol.107.5.1243
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发表时间：
1971
影响因子：
4.4
通讯作者：
D. Bing
中科院分区：
文献类型：
--
作者：
D. Bing
人补体的第一组分C1 β已通过亲和色谱法在由共价连接至人IgG的琼脂糖组成的树脂上纯化。该树脂选择性吸附在最终离子强度为0.03和pH为6.4下制备的人血清沉淀物中可检测到的至少90%的Cl-。用0.2M 1，4-二氨基丁烷从树脂上洗脱C1-C12。几乎所有的蛋白质都被回收，并且无论用N-苄氧羰基-L-酪氨酸-对硝基苯基酯还是用EAC 4、C2和C-EDTA测定，C1 β都具有高比活性。通过该方法获得的蛋白质确实是C1 β的证据是基于C1 β抑制剂抑制酶活性的能力、抗C1 β抗血清抑制EAC 4 β，2 β形成的能力和洗脱的蛋白质形成EAC 1 β，4中间体的能力。该方法似乎是制备大量人C1 β的简单可靠的技术。
The first component of human complement, C1̄, has been purified by affinity chromatography on a resin consisting of Sepharose covalently linked to human IgG. This resin selectively adsorbs at least 90% of the C1̄ detectable in a precipitate of human serum prepared at a final ionic strength of 0.03 and pH of 6.4. The C1̄ is eluted from the resin with 0.2 M 1, 4-diaminobutane. Virtually all the protein is recovered and the C1̄ has a high specific activity whether assayed with N-carbobenzoxy-L-Tyrosine-para-nitrophenyl ester or with EAC4, C2 and C-EDTA. Evidence that the protein obtained by this procedure is indeed C1̄ is based on the ability of C1̄ inhibitor to inhibit the enzymatic activity, of anti-C1s̄ antiserum to inhibit EAC4̄,2̄ formation and of the eluted protein to form the EAC1̄,4 intermediate. The procedure appears to be a simple reliable technique for preparing large quantities of human C1̄.