抗黄曲霉毒素单克隆抗体F(ab')2片段的制备与活性

摘要

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∶320000,which was 1.5 folds higher than 1Cll.And the sensitivity (IC50) of F (ab')2 fragments was 8.7pg/mL against aflatoxin B1 (AFB1).It concluded that F(ab')2 fragments maintained the sensitivity of the original antibodylC11 but with higher titer.%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1Cll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37C、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∶320000,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.