【摘要】 从分泌抗苏丹红Ⅰ单克隆抗体(mAb)的杂交瘤细胞株SU211中克隆抗体轻链Lc基因和重链Fd基因,将Lc、Fd基因以及Linker克隆到载体pET24a(+)中,构建scFab表达载体并在E.coli BL21(DE3)获得表达,SDSPAGE和Westernblot分析表明其表达形式为包涵体,相对分子质量约为57 kD,与理论预期值相符。经复性后,ELISA鉴定结果表明复性抗体对苏丹红Ⅰ具有很高的结合特异性。本研究为新型基因工程抗体的构建以及苏丹红新型免疫检测技术的建立奠定基础。更多还原

【Abstract】 Genes encoding light chain(Lc) and Fd fragment of monoclonal antibody(mAb) against Sudan I were cloned from cell line SU211,and constructed into a single chain Fab antibody(scFab) with a linker.Then the scFab was expressed in E.coli BL21(DE3) with the expression vector pET24a(+ ).The result in SDS PAGE and Western blotting indicated the scFab was expressed in the form of inclusion body,with a relative molecular weight of 57 kD.After renatualization,high degree of specificity of scFab binding to Sudan I could be demonstrated by ELISA.The result of the present study lays a potential foundation for the construction of new genetic engineering antibody and the establishment of new method to detect the antibodies against Sudan I.更多还原