论文部分内容阅读
目的制备适合放免显像的抗癌胚抗原微型抗体。 方法在已获得的抗癌胚抗原重链可变区 (VH)及轻链可变区 (VL)基因的基础上将重链可变区 (VH)与轻链可变区 (VL)基因直接连接制成微型抗体基因 ,并在大肠杆菌进行表达。 结果大肠杆菌高效表达了微型抗体C端 6×His的融合蛋白 ,表达量达菌体总蛋白的 15 % ,SDS -PAGE和Western印迹显示表达物分子量为 2 6kd与基因编码蛋白的理论推算值相符 ,RIA表明表达物与CEA特异性结合。结论大肠杆菌表达是制备抗癌胚抗原微型抗体的有效途径
Objective To prepare anti-carcinoembryonic antigen microantibodies suitable for radioimmunoimaging. Methods Based on the obtained VH and VL genes of anti-CEA, the VH and VL genes were directly amplified Linked to make mini-antibody gene, and expressed in E. coli. Results Escherichia coli expressed a 6 × His fusion protein at the C-terminus of the miniature antibody with an expressed level of 15% of the total bacterial proteins. SDS-PAGE and Western blotting showed that the expressed protein had a molecular weight of 26 kd which was consistent with the theoretical estimate of the gene encoding protein , RIA indicates that the expression specifically binds to CEA. Conclusion Escherichia coli expression is an effective way to prepare anti-carcinoembryonic antigen microantibodies