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单克隆抗体亲和层析法纯化重组溶葡萄球菌酶
引用本文:黄晋江,吴宏宇,张继恩,黄青山. 单克隆抗体亲和层析法纯化重组溶葡萄球菌酶[J]. 生物工程学报, 2009, 25(1): 0147-0151
作者姓名:黄晋江  吴宏宇  张继恩  黄青山
作者单位:1. 上海高科联合生物技术研发有限公司,上海,201206
2. 复旦大学生命科学学院遗传学研究所,遗传工程国家重点实验室,上海,200433
摘    要:溶葡萄球菌酶能够特异性杀灭金黄色葡萄球菌且不易产生耐药性, 有望成为治疗葡萄球菌属细菌引发感染的特效药物。为获得高纯度的重组溶葡萄球菌酶以达到药用标准, 本研究构建了一种以重组溶葡萄球菌酶单克隆抗体为配体的亲和层析纯化方法。纯化后的重组溶葡萄球菌酶纯度大于95%, 得率大于90%, 即使重复使用30多次, 纯化效率不变。且经比色法鉴定纯化后的重组溶葡萄球菌酶仍具有良好的活性。该方法步骤简单, 纯化效果好, 为生产高纯度重组溶葡萄球菌酶奠定了基础。

关 键 词:溶葡萄球菌酶  单克隆抗体  亲和层析
收稿时间:2008-06-20

Purification of recombinant lysostaphin by monoclonal antibody affinity chromatography
Jinjiang Huang,Hongyu Wu,Jien Zhang and Qingshan Huang. Purification of recombinant lysostaphin by monoclonal antibody affinity chromatography[J]. Chinese journal of biotechnology, 2009, 25(1): 0147-0151
Authors:Jinjiang Huang  Hongyu Wu  Jien Zhang  Qingshan Huang
Affiliation:Shanghai Hi-tech United Bio-technological Research & Development Co. Ltd, Shanghai 201206, China;Shanghai Hi-tech United Bio-technological Research & Development Co. Ltd, Shanghai 201206, China;Shanghai Hi-tech United Bio-technological Research & Development Co. Ltd, Shanghai 201206, China;State Key Laboratory of Genetic Engineering, Insititute of Genetics, School of Life Science, Fudan University, Shanghai 200433, China
Abstract:Lysostaphin, a specific endopeptidase enzyme derived from Staphylococcus aureus, is a bactericidal agent against Staphylococcus and difficult to be drug-resistant. This study established monoclonal antibody affinity chromatography to obtain lysostaphin of high purity for drug-use standard. The purified Lysostaphin was of > 95% purity and its recovery rate more than 90%. Moreover, the affinity column kept its efficiency of purification invariable after more than 30 times repeat. Also, the dye release assay validated that the purified lysostaphin had significant bactericidal activity. This method was simple and of high efficacy for the lysostaphin purification and showed its potency in commercial production.
Keywords:lysostaphin   monoclonal antibody   affinity chromatography
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