噬菌体展示技术制备甲氧基有机磷农药抗独特型抗体

Preparation of Anti-idiotype Antibodies of O,O-Dimethyl Organophosphorus Pesticides by Phage Display Technology

抗独特性抗体可用于建立针对农药等小分子物质的非竞争免疫检测模式。本研究将甲氧基有机磷农药通用半抗原(S-羧甲基O-,O-二甲基二硫代磷酸酯,CMP)连接至琼脂糖凝胶,进而对甲氧基有机磷农药广谱特异性抗体进行纯化;以纯化抗体为固相抗原,以人源scFv抗体库(Tomlinson I+J)为抗体源,进行抗独特型抗体的筛选;利用所筛选到的抗独特型抗体建立非竞争免疫检测技术。经过鉴定,共筛选出12个阳性克隆,其中D11和B9分别属于β型和α型抗独特型抗体。利用D11和B9建立的非竞争免疫分析法对马拉硫磷的IC50为(113.7±34.2)μg/L,检出限为10.5μg/L。

Anti-idiotype antibody（AId or Ab2） can be applied to develop noncompetitive immuno-assay for hapten molecules.In this study,hapten molecules were immobilized on sepharose,and then used for antibody purification.Affinity purified positive and negative primary antibodies were used for bio-panning of anti-idiotype antibody from phage display antibody library.After three round of panning,3α-AId and 9β-AId clones were achieved.Clones D11 and B9,identified as β-AId and α-AId respectively,were selected for developing a noncompetitive immunoassay system.The IC50 value and detection limit of this method for malathion were calculated to be（113.7±34.2） μg/L and 10.5 μg/L,respectively.