结核分枝杆菌ESAT-6抗原鼠单克隆抗体的制备及初步鉴定

摘要

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 IgGl/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.%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IgGl/K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.