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高特异性黄曲霉毒素B_1单克隆抗体的制备及特性研究
引用本文:肖智,李培武,张奇,张文,丁小霞.高特异性黄曲霉毒素B_1单克隆抗体的制备及特性研究[J].中国油料,2011(1):66-70.
作者姓名:肖智  李培武  张奇  张文  丁小霞
作者单位:[1]中国农业科学院油料作物研究所,湖北武汉430062 [2]农业部油料作物生物学重点开放实验室,湖北武汉430062 [3]农业部油料及制品质量监督检验测试中心,湖北武汉430062
基金项目:农业部948项目(2009-Z46 2010-G1); 国家油菜现代产业技术体系(nycytx-005); 国家科技支撑计划项目(2010BAD01B07 2011BAD02D02); 中国农业科学院油料作物研究所中央级公益性科研院所基本科研业务费专项
摘    要:本研究将黄曲霉毒素B1转化为半缩醛B2a,在硼氢化钠(NaBH4)还原作用下与载体蛋白偶联制备完全抗原。将制备的完全抗原免疫Balb/c小鼠,经4次免疫后取其脾脏与小鼠骨髓瘤细胞Sp2/0细胞融合,采用半固体培养基筛选后鉴定,获得杂交瘤细胞株3A12,抗体的灵敏度可达6.1±0.025ng/mL,抗体与其它黄曲霉毒素B2、G1及G2的交叉反应率依次为7.8%、20.2%及0.6%,与黄曲霉毒素M1交叉反应率小于0.1%。本研究为研发花生等农产品黄曲霉毒素B1特异性免疫分析技术及产品奠定了重要基础。

关 键 词:黄曲霉毒素B1  高特异性单克隆抗体  特性

Production and characteristics of specialised monoclonal antibodies against aflatoxin B_1
Authors:XIAO Zhi  LI Pei-wu  ZHANG Qi  ZHANG Wen  DING Xiao-xia
Affiliation:1.Oil Crops Research Institute of Chinese Academy of Agriculture Sciences,Wuhan 430062,China; 2.Key Laboratory of Oil Crops Biology of the Ministry of Agriculture,Wuhan 430062,China; 3.Quality Inspection and Test Center for Oilseeds Product,MOA,Wuhan 430062,China)
Abstract:Aflatoxins are important mycotoxins of contaminated peanut products.Aflatoxin B1 is the most toxic,accounting for over 70% of aflatoxin analogues.In present paper,aflatoxin B1 was transformed into hemiacetal,namely aflatoxin B2a,and then conjugated to protein as immunogen by reduction of sodium borohydride.After the fourth boost immunization,spleen cells of the immunized Balb/c mouse were fused with freshly isolated SP2/0 myeloma cells.Through selecting and identifying with enzyme-linked immunosorbent assay(ELISA),one clone 3A12 was obtained from cultured semi-solid medium.Sensitivity of antibody 3A12 reached 6.1±0.025ng/mL.The cross-reactivities with other aflatoxin analogues(aflatoxin B2,G1 and G2) were 7.8%,20.2% and 0.6% respectively,with aflatoxin M1 less than 0.1%.It offered an important foundation for aflatoxin B1-specific immuno-assay technologies and products.
Keywords:Aflatoxin B1  High specificity monoclonal antibodies  Identification
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