抗Cry1c蛋白兔单克隆抗体的制备和鉴定

Preparation and Identification of Anti-Cry1c Protein Rabbit Monoclonal Antibodies

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。

Crylc protein was expressed by prokaryotic plasmid and after purification progress, Crylc protein was injected into rabbits as an antigen. After serum titer detection, cells fusion, hybridoma cell screening, plasmid DNA re- combination, anti-Crylc monoelonal antibodies（McAb） were obtained. After detecting some relation characters by ELISA, the results showed that the titer of McAb surpass 3.2×10^4, the affinity constant was about 2.4×10^9, there was little cross-reaction between CrylAe and anti-Crylc McAb, and no obvious cross-reaction between CrylAb and anti-Crylc McAb. The embedding amount of Cryle proteic was 400 ng/mL detected by indirect ELISA. It may be decide as positive.