结核分枝杆菌ESAT-6抗原鼠单克隆抗体的制备及初步鉴定  

Preparation and Characterization of Murine Monoclonal Antibodies Against ESAT-6 Antigen of Mycobacterium tuberculosis

作　　者：刘树玲[1] 杨秀旭[1] 李浩[1] 任军[1] 李建民[1] 付玲[1] 徐俊杰[1] 陈薇[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《生物技术通讯》2011年第2期258-260,共3页Letters in Biotechnology

基　　金：国家自然科学基金(30972772)

摘　　要：目的:制备重组结核分枝杆菌ESAT-6抗原鼠单克隆抗体。方法:采用杂交瘤技术,获得了12株针对结核分枝杆菌ESAT-6抗原的鼠单克隆抗体杂交瘤细胞株,对其中的5株进行了小鼠腹水的制备及相关鉴定。结果:5株单克隆抗体的腹水效价达到1:512 000～1:1 024 000,纯化后纯度高于90%,抗体亚类(型)均为IgG1/κ型,ELISA结果显示制备的单克隆抗体与结核分枝杆菌ESAT-6抗原可发生特异反应。结论:制备了结核分枝杆菌ESAT-6抗原鼠单克隆抗体,为结核分枝杆菌ESAT-6的生物学功能研究奠定了基础。Objective：To produce murine monoclonal antibodies（mAbs） against recombinant ESAT-6 antigen of Mycobacterium tuberculosis.Methods：Twelve stably secreting anti-ESAT-6 specific mAbs cell strains were obtained with hybridoma cell fusion technique.The five strains of above cell strains were identified and used to prepare the ascite fluids.Results：The ELISA titers in the ascite fluids of the five mAbs were 1：512 000～1：1 024 000.The ascite mAbs were purified up to 90% purity.IgG isotypes showed that all the mAbs secreted by the five stains belong to IgG1/κ subclass,ELISA proved that the five mAbs reacted specifically with ESAT-6 antigen of M.tuberculosis.Conclusion：The specific anti-ESAT-6 mAbs were prepared,which may lay a foundation for studying biological functions of ESAT-6.

关 键 词：结核分枝杆菌 ESAT-6 杂交瘤 单克隆抗体 

分 类 号：Q813.2[生物学—生物工程]