磁珠标记的氯霉素直接竞争ELISA方法的建立  

Establishment of cdELISA marked by immunomagnctic beads for the determination of chloramphenicol

作　　者：李因来[1] 陈正贤[2] 高乃波[1] 施曼玲[1] 

机构地区：[1]杭州师范大学生命与环境科学学院,浙江杭州310036 [2]浙江大学生物技术研究所,浙江杭州310029

出　　处：《中国兽医学报》2010年第4期498-501,共4页Chinese Journal of Veterinary Science

基　　金：杭州市科技局市属高校重点实验室科技创新项目(2006831H07)

摘　　要：采用本实验室制备的氯霉素(CAP)单克隆抗体,以包被抗原1∶6 000和酶标CAP单抗1∶8 000的稀释比例为工作浓度,建立CAP直接竞争ELISA(cdELISA)方法,cdELISA检测灵敏度达到0.1μg/L。利用xMag异硫氰酸根末端磁粒标记CAP单克隆抗体制备免疫磁珠,用该免疫磁珠富集样品中的CAP。经过乙酸乙酯抽提免疫磁珠表面吸附的CAP,再通过cdELISA检测,可使CAP的检测灵敏度提高50倍,即达到0.002μg/L。A direct competitive enzyme-linked immunosorbent assay（cdELISA） was developed for the determination of chloramphenicol（CAP）,with coating antigen and horseradish peroxidase（HRP）-labeled conjugated monoclonal antibody（McAb） against CAP at 1∶6 000 and 1∶8 000 dilution,respectively.The detection limit for CAP was 0.1 μg/L in cdELISA.Immunomagnetic beads（IMB） were prepared by xMag-magnetic beads marking McAb against CAP.CAP was enriched from the samples through adsorption of IMB,and CAP which was extracted from surface of IBM by ethylacetate was detected by cdELISA.With above method,the limit of CAP detection is improved 50 times and to be 0.002 μg/L in cdELISA.

关 键 词：氯霉素 单克隆抗体 ELISA 免疫磁珠 

分 类 号：S852.43[农业科学—基础兽医学]