伏马菌素B_1单克隆抗体制备及化学发光免疫检测方法的建立  

Production of Monoclonal Antibodies for Detection of Fumonisin B1 by Chemiluminescence Enzyme-linked Immunosorbent Assaay

作　　者：许杨[1] 邹龙[1] 刘师文[2] 刘京[1] 陈波[1] 

机构地区：[1]南昌大学食品科学与技术国家重点实验室,中德联合研究院,南昌330047 [2]江西省疾病预防控制中心,南昌330029

出　　处：《分析化学》2012年第11期1735-1739,共5页Chinese Journal of Analytical Chemistry

基　　金：国家"863"计划资助项目(No.2007AA10Z427)

摘　　要：制备了高亲和力的伏马菌素B_1(FB_1)单克隆抗体,进而建立了谷物中FB_1的直接竞争化学发光酶联免疫检测方法(dc-CLEIA)。采用碳化二亚胺法成功合成了FB_1人工抗原,经杂交瘤技术获得分泌FB_1特异性抗体的细胞株,高碘酸钠法酶标单克隆抗体后,建立了FB_1的dc-CLEIA,本方法的IC_(50)值为1.43μg/L,线性范围为0.32～8.40μg/L,检出限为0.13μg/L,竞争反应时间仅需20 min。玉米样品加标回收率为100.2%～115.4%,相对标准偏差小于8.7%,谷物样品的检测结果与高效液相色谱法及商业化ELISA试剂盒检测结果相符。Based on monoclonal antibodies with high affinity against fumonisin B1,a direct competitive chemiluminescence enzyme-linked immunosorbent assay（dc-CLEIA） for detection of FB, in cereal was developed.FB]artificial antigens were synthesized by the carbodiimide crosslinking methods,and then a hybridoma cell that secreted the antibody selectively identifies FB） was selected through cell fusion.The monoclonal antibody titer was found as higher as 1.28×10~6,which was labeled with horse radish peroxidase（HRP） through sodium periodate activation for development of dc-CELIA.A good linearity was achieved over a concentration range of 0.32—8.40μg/L,the IC50 value and the limit of detection were 1.43μg/L and 0.13μg/L respectively.Meanwhile,the competitive reaction time was only 20 min.The recoveries of FB_1 in all tissues were between 100.2%—115.4%and the relative standard deviation was less than 8.7%.The dc-CLEIA was applied in quantification of FB,elimination in cereal,the results showed good correlations of the results obtained by HPLC and a commercial ELISA kit.

关 键 词：伏马菌素B_1 单克隆抗体 化学发光酶联免疫分析 谷物 

分 类 号：TS210.7[轻工技术与工程—粮食、油脂及植物蛋白工程] O657.3[轻工技术与工程—食品科学与工程]