醋酸甲羟孕酮(MPA)人工抗原的合成与单克隆抗体的制备

为研制检测醋酸甲羟孕酮（MPA）的免疫学检测技术进行MPA单克隆抗体的制备与鉴定，研究分别采用碳二亚胺法、混合酸酐法将MPA偶联到牛血清白蛋白（BSA）和鸡卵清白蛋白（OVA）上制备免疫原（MPA—BSA）与包被原聊PA—OVA），通过BALB／c小鼠免疫、细胞融合、间接ELISA和阻断ELISA筛选、有限稀释法克隆亚克隆获得分泌MPA单克隆抗体的杂交瘤细胞株，并通过ELISA方法对其抗体效价、特异性、敏感性、亚类及稳定性进行鉴定。结果表明：成功合成2种人工抗原，MPA与BSA、OVA的偶联比分别为8：1、12：1；获得1株稳定分泌MPA单克隆抗体（McAb）的杂交瘤细胞株2G4H9，McAb腹水的间接ELISA效价为1：8×10^4，IgG1亚类，K型轻链，腹水抗体IC50为5．0ng·mL^-1，与甲地孕酮、盐酸克伦特罗、莱克多巴胺和己烯雌酚交叉反应率为12．5％、〈0．01％、〈0．01％和〈0．01％。作者认为：MPA单克隆抗体特异性和敏感性良好，杂交瘤细胞株性能稳定，为进一步研究MPA相关免疫学分析技术奠定了基础。

Preparation of Artificial Antigen for Medroxyprogesterone Acetate(MPA) and McAb Against MPA

To prepare monoclonal antibody against medroxyprogesterone acetate(MPA) for the development of immunological analysis methods for MPA residues detection.The hapten MPA was coupled to bovine serum albumin(BSA) and ovalbulmin(OVA) by using di- isopropylcarbodiimide method and mixed anhydrides method to synthesize artificial immunogen and coating antigen,respectively,and the artificial antigen MPA-BSA,MPA-OVA were confirmed by UV spectrum and SDS-PAGE;then the monoclonal antibodies(McAb) against MPA was prepar...