氯霉素人工抗原的合成及多克隆抗体的制备  

Synthesis of Artificial Antigen of Chloramphicol and Preparation of Polyclonal Antibody

作　　者：陆蕾[1] 刘娜[1] 倪庚[1] 王琳[1] 郑晓冬[1] 

机构地区：[1]浙江大学生物系统工程与食品科学学院,杭州310029

出　　处：《中国食品学报》2011年第2期177-184,共8页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家高技术研究发展计划项目(No.2007AA10Z436)

摘　　要：采用重氮化法合成氯霉素人工抗原。通过紫外扫描、凝胶电泳、基质辅助激光解吸飞行时间质谱(MAL-DI-TOFMS)鉴定人工抗原。用该人工抗原免疫兔子,获得多抗血清。采用酶联免疫法测定抗体效价,并对ELISA条件进行初步优化。选择与氯霉素结构相似的4种药物进行交叉反应,计算交叉反应率。结果表明免疫抗原偶联率为1∶22,最佳包被抗原质量浓度为0.25μg/mL,抗体效价128000。优化后,通过间接竞争ELISA法建立标准曲线,IC50为0.26μg/mL,4种药物与抗体的交叉反应率均小于0.1%,特异性显著。Artificial antigen of chloramphenicol was synthesized by diazotization method.The artificial antigen were identified by UV spectrophotometer,SDS-PAGE electrophoresis and MALDI-TOFMS.Rabbits were immunized with that artificial antigens.The polyantiserum was obtained,and the antiserum titre was determined by ELSIA.The condition of ELISA was optimized.Four antibiotics were chosed to calculate the cross reactivity because of their smilar structure.The result showed conjugation ratio was 22∶1 and working concentration of coating antigen was 0.25 μg/mL.The standard curve was eatablished and the value of IC50 was 0.26 μg/mL.Antiserum titre reached 128 000.The cross reactivity of 4 drugs and antibody were less than 0.1% with high specificity.

关 键 词：氯霉素 人工抗原 重氮化 优化 

分 类 号：S859.84[农业科学—临床兽医学]