抗黄曲霉毒素单克隆抗体F(ab')2片段的制备与活性  

Preparation and activity of F(ab′)2 fragments of monoclonal antibody against aflatoxin

作　　者：雷佳文[1,2,3,4] 李培武[1,2,3,5,4] 张奇[1,2] 张兆威[1,2] 丁小霞[1,5] 张文[1,4] 

机构地区：[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与遗传育种重点实验室,湖北武汉430062 [3]农业部生物毒素检测重点实验室,湖北武汉430062 [4]农业部油料及制品质量监督检验测试中心,湖北武汉430062 [5]农业部油料产品质量安全风险评估实验室,湖北武汉430062

出　　处：《中国油料作物学报》2013年第3期317-320,共4页Chinese Journal of Oil Crop Sciences

基　　金：农业部948重点项目(2011-G5;2010-G1);国家科技支撑计划(2012BAB19B09;2012BAK08B03);国家自然科学基金(31171702;31101299);公益性农业行业专项(201203094;201203037);现代农业产业技术体系建设专项(CARS-13);中国农业科学院所长基金(1610172010003);湖北省自然科学基金(2011CDB358)

摘　　要：为剧毒靶标(黄曲霉毒素)绿色分析提供高效抗体,在已有抗黄曲霉毒素杂交瘤细胞株1C11的基础上,通过小鼠腹水法制备单克隆抗体,经胃蛋白酶酶解,制备F(ab')2片段,发现在优化条件37℃、pH4.1柠檬酸缓冲液中酶解4.5h,可高效制备F(ab')2片段。通过酶联免疫吸附法(ELISA)比较了抗体片段与原始抗体的识别活性,发现F(ab')2片段效价达到1∶320 000,是原抗体效价的1.5倍;灵敏度(IC50)为8.7pg/mL,保持了原抗体的抗原结合能力。To provide high effect materials for anti-idiotype and anti-immune complex antibodies in aflatoxin research,monoclonal antibody 1C11 from mouse ascites based on hybridoma cell 1C11 was used.The antibody was prepared and digested by pepsin at 37℃ in pH4.1 citrate buffer for 4.5h for the preparation of F（ab′）2 fragments.After purification,the characteristics of F（ab′）2 fragment were identified by ELISA.Results showed that the titer of F（ab′）2 fragments was 1∶320 000,which was 1.5 folds higher than 1C11.And the sensitivity（IC_50） of F（ab′）2 fragments was 8.7pg/mL against aflatoxin B_1（AFB_1）.It concluded that F（ab′）2 fragments maintained the sensitivity of the original antibody1C11 but with higher titer.

关 键 词：黄曲霉毒素 单克隆抗体 F(ab')2片段 活性 

分 类 号：S379.7[农业科学—农产品加工]