胸腺肽α_1单克隆抗体的制备及其鉴定

Preparation and identification of monoclonal antibody against thymosin alpha 1

目的 研制胸腺肽α1(Tα1)单克隆抗体 ,为Tα1作用机制的阐明奠定基础。方法 用戊二醛将Tα1和牛血清白蛋白进行交联 ,以其交联物为抗原 ,按常规淋巴细胞杂交瘤技术制备单克隆抗体 ,并采用间接酶联免疫吸附 (ELISA)法和Western免疫印迹来鉴定单克隆抗体的特异性。结果 建立了分泌抗Tα1抗体的杂交瘤细胞株 ,能特异性地与Tα1结合 ,但与单纯牛血清白蛋白、酵母抽提物和转染Tα1酵母未诱导表达产物无反应。其抗体效价为 1∶10 2 4 0 ,Ig亚类为IgG2b。结论 本研究建立了针对Tα1高效价、高特异性单克隆抗体细胞株。

Objective To produce monoclonal antibody (McAb) against thymosin alpha 1 and make a foundation for the research and identification of thymosin alpha 1. Methods McAb against thymosin alpha 1 was prepared by the lymphocyte hybridoma technique, the specificity of McAb was identified by indirect ELISA and Western blotting method. Results The hybridoma producing antibody against thymosin alpha 1 was obtained, the titer of McAb was 1∶10 240, the subtype of Ig was IgG 2b , the McAb could react to thymosin alpha 1,but failed to react to other protein such as bovine serum albumin and yeast extract. Conclution It has been established that a hybridoma cell line can produce high titer and specificity monlclonal antibody against thymosin alpha 1.