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学科主题临床医学
A method of purifying intact complement factor H from human plasma
Wang, Feng-Mei; Yu, Feng; Zhao, Ming-Hui
关键词Complement factor H Human plasma Purification Chromatography
刊名PROTEIN EXPRESSION AND PURIFICATION
2013-10-01
DOI10.1016/j.pep.2013.07.014
91期:2页:105-111
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
研究领域[WOS]Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]HEMOLYTIC-UREMIC SYNDROME ; MACULAR DEGENERATION ; FLUID-PHASE ; DISEASE ; PROTEIN ; PURIFICATION ; RECOGNITION ; DEFICIENCY ; SYSTEM ; SERUM
英文摘要

The aim of this study was to establish a method of purifying intact complement factor H (CFH) from human plasma. CFH was isolated from human plasma by polyethylene glycol (PEG) precipitation, following three sequential chromatographic columns, which consisted of L-lysine Sepharose column, Resource Q column and Sephacryl S-300 High Resolution HiPrep 16/60 column. All the above steps were performed at 4 degrees C by Fast Protein Liquid Chromatography (FPLC) AKTA Purifier 10 with Frac-900. Identification of the purified CFH was confirmed by SOS-PAGE and Western blot. The following functions of the purified CFH were further analyzed compared with the commercial CFH in vitro: (1) binding ability with C3b; (2) binding ability with mCRP; (3) the protecting function of the hemolysis of sheep red blood cells; (4) the cofactor role for complement factor I-mediated proteolytic inactivation of C3b. Homogeneous CFH was purified from the plasma fraction through the above four steps. The purity and the functions of the purified CFH were comparable to the commercial CFH. The yield of CFH was 26 +/- 3% in our study. Compared with previous methods, our method was high yield with high purity. We established a stable and feasible system for purifying intact CFH, which could be used in the lab and clinical investigations. (C) 2013 Elsevier Inc. All rights reserved.

语种英语
WOS记录号WOS:000324607100001
项目编号2012CB517702 ; 81021004 ; 81100497 ; 7102150
资助机构Chinese 973 project ; National Natural Science Foundation of China ; Beijing Natural Science Foundation
引用统计
被引频次:5[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/65054
专题北京大学第一临床医学院_肾脏内科
作者单位1.Peking Univ, Hosp 1, Dept Med, Div Renal, Beijing 10034, Peoples R China
2.Key Lab Chron Kidney Dis Prevent & Treatment, Minist Educ China, Beijing 100034, Peoples R China
3.Peking Univ, Inst Nephrol, Beijing 10034, Peoples R China
4.Key Lab Renal Dis, Minist Hlth China, Beijing 10034, Peoples R China
推荐引用方式
GB/T 7714
Wang, Feng-Mei,Yu, Feng,Zhao, Ming-Hui. A method of purifying intact complement factor H from human plasma[J]. PROTEIN EXPRESSION AND PURIFICATION,2013,91(2):105-111.
APA Wang, Feng-Mei,Yu, Feng,&Zhao, Ming-Hui.(2013).A method of purifying intact complement factor H from human plasma.PROTEIN EXPRESSION AND PURIFICATION,91(2),105-111.
MLA Wang, Feng-Mei,et al."A method of purifying intact complement factor H from human plasma".PROTEIN EXPRESSION AND PURIFICATION 91.2(2013):105-111.
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