论文部分内容阅读
为获得能够用于夹心ELISA检测的配对抗CP4-EPSPS蛋白单克隆抗体,构建CP4-EPSPS原核表达载体并转化大肠杆菌Rossetta,获得高效表达。通过对可溶性蛋白纯化,获得了高纯度目的蛋白。以纯化后的CP4-EPSPS蛋白作为抗原免疫BALB/c小鼠,经过细胞融合和筛选获得2株抗CP4-EPSPS蛋白单克隆抗体(1A5和8A3)。经鉴定,这2株抗体可以有效地识别高温变性和天然CP4-EPSPS蛋白;叠加ELISA分析表明两株抗体识别的抗原表位不同,说明两株抗体能够用于夹心ELISA检测CP4-EPSPS蛋白。
To obtain a paired anti-CP4-EPSPS protein monoclonal antibody that can be used for sandwich ELISA detection, a prokaryotic expression vector for CP4-EPSPS was constructed and transformed into E. coli Rossetta for high expression. By purifying the soluble protein, high purity target protein was obtained. BALB / c mice were immunized with the purified CP4-EPSPS protein as antigen. Two anti-CP4-EPSPS monoclonal antibodies (1A5 and 8A3) were obtained by cell fusion and screening. The two antibodies were identified as high temperature denatured and native CP4-EPSPS protein. The superposition ELISA showed that the two antibodies recognized different antigen epitopes, indicating that the two antibodies could be used to detect CP4-EPSPS protein by sandwich ELISA.