庆大霉素单克隆抗体的制备及ELISA检测方法研究  

Preparation of monoclonal antibody against Gentamicin and development of ELISA for detection of Gentamicin residues

作　　者：金涌[1] 陈立本[1,2] 邹明强[1] 郝钢[1] 王羚鸿[1] 田世民[1] 马吉湘[1] 

机构地区：[1]中国检验检疫科学研究院,北京朝阳100123 [2]中国农业大学动物医学院,北京海淀100193

出　　处：《中国兽医杂志》2009年第8期9-12,共4页Chinese Journal of Veterinary Medicine

基　　金："十一五"国家科技支撑计划重大项目(2006BAK10B09);中国检验检疫科学研究院基本科研业务费专项资金资助项目(2007JK009)

摘　　要：本研究采用碳二亚胺法将庆大霉素与载体蛋白偶联形成完全抗原，用基质辅助激光解吸／电离质谱进行了表征。以人工抗原免疫BALB／C小鼠，通过杂交瘤技术获得了分泌抗庆大霉素单克隆抗体并能稳定传代的杂交瘤细胞株，建立了庆大霉素间接竞争ELISA检测方法，其线性检测范围为0．1～5ng／mL，半数抑制浓度（IC50）为0．8ng／mL，与其他化合物的交叉反应率小于0．01％。按药典剂量给患乳房炎奶牛肌肉注射庆大霉素注射液，不同时间段采集牛奶样品，按本研究建立的竞争ELISA方法测定了牛奶中的庆大霉素残留量。An ester activation method was employed to synthesize Gentamicin antigens using KLH and BSA as carrier proteins. The conjugate was identified by matrix-assisted laser desorption/ionization timeof-flight mass spectra （MALDI-TOF MS）. Monoclonal antibody against Gentamicin was prepared, and a competitive ELISA was developed using the monoclonal antibody to detect Gentamicin. Based on the calibration curve, the linear detection was 0.1 to 5 ng/mL and the IC50 was 0.8 ng/mL. No cross-reactivity of the antibody with other compounds was observed, indicating that the antibody is highly specific for Gentamicin. Using competitive ELISA method developed in this study, Gentamicin concentrations were successfully monitored in milk after intramuscular administration of Gentamicin.

关 键 词：庆大霉素 基质辅助激光解吸/电离质谱法 单克隆抗体 竞争ELISA 牛奶样品 

分 类 号：S859.796[农业科学—临床兽医学] S852.43[农业科学—兽医学]