单克隆抗体亲和层析法纯化重组溶葡萄球菌酶

Purification of recombinant lysostaphin by monoclonal antibody affinity chromatography

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。

Lysostaphin, a specific endopeptidase enzyme derived from Staphylococcus aureus, is a bactericidal agent against Staphylococcus and difficult to be drug-resistant. This study established the 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.