A RAPID FPLC METHOD FOR PURIFICATION OF THE 3RD COMPONENT OF HUMAN AND GUINEA-PIG COMPLEMENT

被引：21

作者：

BASTA, M

HAMMER, CH

机构：

[1] Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Disease, National Institutes of Health, Bethesda, MD

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1991年 / 142卷 / 01期

关键词：

CHROMATOGRAPHY; FAST PROTEIN LIQUID; ANION-EXCHANGE; C3; HUMAN; GUINEA PIG; PURIFICATION METHOD;

D O I：

10.1016/0022-1759(91)90290-V

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method is described for the purification of human and guinea pig C3 from small amounts of serum. This procedure requires only two steps - polyethylene glycol (PEG) precipitation and fast protein liquid chromatography (FPLC) Mono Q HR 10/10 ion exchange chromatography. The protocol takes less than two hours to complete and yields 4-6 mg of purified C3. Similar results, in terms of antigenic and functional recovery, were obtained for both human and guinea pig components. About 67% of C3 antigen was recovered from eluted fractions with fully preserved specific activity. Isolated C3 was over 95% pure as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography; this level of purity was confirmed by the absence of any observable contamination as assessed by immunoelectrophoresis using high titer anti-whole human serum. This method allows rapid and reproducible purification of fully active human or guinea pig C3 on a daily basis.

引用

页码：39 / 44

页数：6

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