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通过原核质粒表达重组Cry1c晶体蛋白,将其纯化后作为特定抗原免疫兔子。经多抗血清效价的测定,细胞融合,杂交瘤细胞的筛选,构建重组载体等步骤获得抗Cry1c单克隆抗体。通过ELISA方法测定单克隆抗体相关特性,结果表明,Cry1c兔单克隆抗体效价超过3.2×104,其亲和常数为2.4×109L/mol,该单克隆抗体与Cry1Ac蛋白有微量交叉反应,与Cry1Ab蛋白无明显交叉反应。间接ELISA检测抗原Cry1c蛋白时,当包被量为400ng/mL时,判定为阳性。
Recombinant Cry1c crystal protein was expressed by prokaryotic plasmid, which was purified and used as a specific antigen to immunize rabbits. The anti-Cry1c monoclonal antibody was obtained by the determination of multi-antiserum titer, cell fusion, hybridoma screening, construction of recombinant vector and the like. The results showed that the titer of monoclonal antibody against Cry1c rabbit was over 3.2 × 104, and its affinity constant was 2.4 × 109 L / mol. The monoclonal antibody had a slight cross-reaction with Cry1Ac protein, Protein no significant cross-reaction. Indirect ELISA detection of antigen Cry1c protein, when the coating amount of 400ng / mL, was judged as positive.