脱二氧喹烯酮多克隆抗体的制备及其直接竞争ELISA检测方法的建立  

Preparation of Polyclonal Antibody Against Desoxyquinocetone and Establishment of A Direct Competitive ELISA Method for Desoxyquinocetone Detection

作　　者：曹娟[1] 傅坚英[1] 王延英[1] 张吉斌[1] 

机构地区：[1]华中农业大学生命科学技术学院农业微生物学国家重点实验室,湖北武汉430070

出　　处：《化学与生物工程》2012年第7期81-85,共5页Chemistry & Bioengineering

基　　金：国家973计划资助项目(2009CB118800)

摘　　要：利用脱二氧喹烯酮(DQCT)的酮基为活性基团,分别采用混合酸酐法和碳二亚胺法合成了其免疫原(DQCT-BSA)和包被原(DQCT-OVA),用过碘酸钠氧化法将DQCT-OVA与辣根过氧化物酶(HRP)偶联,制备了酶标抗原(DQCT-OVA-HRP)。用DQCT-BSA免疫Balb/C小鼠,获得了效价1∶128 000的抗DQCT的多克隆抗体。分别用间接竞争ELISA法和直接竞争ELISA法检测DQCT并比较,最终建立了直接竞争ELISA检测方法,其IC50为39.8μg.mL-1,检测范围为1.6～1258.9μg.mL-1。为后续实际样品中脱二氧喹烯酮的检测奠定了基础。Ketone of desoxyquinocetone（DQCT） was used as active group. Immunogen DQCT-BSA and coating antigen DQCT-OVA were synthesized by mixed anhydride method and carbodiimide method, respective- ly. The enzyme labeled antigen DQCT-OVA-HRP was synthesized using oxidize method of NaIO, to conjugate HRP with DQCT-OVA. Polyclonal antibody against DQCT with a titer of 1：128 000 was prepared through im- munizing mouse with DQCT-BSA. DQCT was detected by direct and indirect competitive ELISA methods, re- spectively. A direct competitive ELISA for DQCT detection was established,for which the IC50 was 39.8 μg·mL^-1 and the detection range was 1.6.-1258.9 μg·mL^-1. This study laid a foundation for DQCT detection in real sample.

关 键 词：脱二氧喹烯酮 酶标抗原 竞争酶联免疫吸附分析法 

分 类 号：Q782[生物学—分子生物学] S859.79[农业科学—临床兽医学]