【作者】 范国英； 杨艳艳； 王自良； 邓润广； 张改平；

【Author】 FAN Guoying~1,YANG Yanyan~2,WANG Ziliang~1,DENG Riuguang~2,ZHANG Gaiping~2 (1.The Animal Science College,Henan Institute of Science and Technology,Xinxiang,Henan 453003,China; 2.Institute of Biology and Technique Science,Agricultural Academy of Henan Province,Zhengzhou, 450002,China)

【机构】 河南科技学院动物科学学院； 河南省农业科学院免疫学重点实验室；

【摘要】 用碳二亚胺(DCC)一步法,将经对醛基苯甲酸衍生物CPAMOZ偶联于牛血清白蛋白、卵清蛋白上,合成免疫原CNPAMOZ-BSA、包被原CNPAMOZ-OVA.用紫外线扫描、SDS-PAGE进行鉴定;免疫BaIb/c小鼠,用杂交瘤技术建立分泌NPAMOZ mAb的细胞株;对NPAMOZ mAb的效价、敏感性和特异性进行鉴定.结果表明:CPAMOZ-BSA人工抗原分子结合比为15:1;筛选出分泌NPAMOZ mAb的敏感特异的杂交瘤细胞1株3F9-C4,间接ELISA测定细胞培养上清,效价达到了1:6.4×102,腹水效价为1:1.2×106,对NPAMOZ IC50为0.95μg/L,与其他抗生素交叉反应率低。本试验获得了抗NPAMOZ高价、敏感、特异的mAb,可用于AMOZ的残留检测.更多还原

【Abstract】 The coating antigen CPAMOZ-OVA,immunogen CPAMOZ-BSA of Enzyme marked synthesized with AMOZ was coupled with BSA and OVA by DCC-method.Ultraviolet and SDS-PAGE were used to identify AMOZ artificial antigen. Balb/c mice were immunized by CPAMOZ-BSA,the hybridoma lines that secrete NPAMOZ mAb were established by using monoclonal antibody hybridoma technology.The immunological traits such as titer,sensitivity and specificity of this mAb were characterized.The results showed that the conjugation ratio of NPAMOZ to BSA in NPAMOZ artificial antigen was 15:1.The titer of supernatant hybridoma cell lines of 3F9-C4 was 1:6.4×102 by indirect ELISA.The determination of Ascites effect is 1:1.2×106.IC50 was 0.95μg/L,Cross-reactivity to other compounds is low.The experiment generated AMOZ mAb was high-titer,sensitivity and specificity.This mAb can be used to detecte AMOZ residues in food.更多还原