| C1q、C1r和C1s的快速分离 |
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| 引用本文： | 李文君,陈政良,张丽芸. C1q、C1r和C1s的快速分离[J]. 现代免疫学, 2002, 22(5): 337-339,349 |
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| 作者姓名： | 李文君 陈政良 张丽芸 |
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| 作者单位： | 第一军医大学免疫学教研室,广州,510515 |
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| 基金项目： | 国家自然科学基金资助项目 (No 39970 6 87)，广东省自然科学基金资助项目 (No 0 10 6 0 0 ) |
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| 摘 要： | 建立了一种同时快速分离纯化C1q及酶原形式C1r和C1s的方法。用IgG Sepharose 4B亲和层析将C1q与C1r2 s2 分离 ,接着用DEAE Sephacel离子交换层析将C1r和C1s分离 ,用C1qMcAb Sepharose 4B亲和层析将C1q进一步纯化。在分离Clr和C1s的整个过程中加入蛋白水解抑制剂PMSF和NPGB ,并控制温度在 4℃、pH 6 1、无二价阳离子的条件下 ,得到高度纯化的C1q、C1r和C1s。
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| 关 键 词： | C1q C1r C1s 亲和层析 离子交换层析 |
| 文章编号： | 1001-2478(2002)05-0337-03 |
A Rapid Methed for the Isolation of the C 1q、 C 1r and C1s |
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| LI Wen jun,CHEN Zheng liang,ZHANG Li yun. A Rapid Methed for the Isolation of the C 1q、 C 1r and C1s[J]. Current Immunology, 2002, 22(5): 337-339,349 |
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| Authors: | LI Wen jun CHEN Zheng liang ZHANG Li yun |
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| Abstract: | It has established an isolation procedure for C1q,C1r and C1s,which was employed IgG Sepharose 4B affinity chromatography followed by DEAE Sephacel ion exchange chromatography and C1q McAb Sepharose 4B affinity chromatography All steps were performed at low temperature 4℃ and two proteolytic inhibitors,PMSF and NPGB,were added during affinity chromatography on IgG Sepharose 4B The novel condition was to keep the pH at 6 1 during the entire procedure,where activation was markedly depressed In addition,purification was improved by washing the IgG Sepharose 4B column with a buffer free of divalent cations immediately prior to elution of the C1r and C1s with EDTA Little or no activated material was detected in these highly purified fractions |
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| Keywords: | C1q C1r C1s affinity chromatography ion exchange chromatography |
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