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在人乳腺癌和其他肿瘤细胞表面通常过表达一 2 8ku膜蛋白 ,可作为肿瘤导向治疗的作用靶 .利用基因工程技术 ,将 6C6单链抗体 (ScFv6C6)同缺失细胞结合区的绿脓杆菌外毒素A(PE40 )相连 ,构建成 6C6免疫毒素 (ScFv6C6_PE40 ) ,其中ScFv6C6由能特异识别 2 8ku蛋白的 6C6单克隆抗体的重链可变区和轻链可变区连接而成 . 6C6免疫毒素的原核表达水平为 3 3 % ,约 5 5mg/L菌液 .利用HisTrap(镍离子螯合 )柱层析纯化 ,得到纯度为 3 3 2 %的 6C6免疫毒素 ,其可识别MDA 2 3 1人乳腺癌细胞表面上的肿瘤相关抗原 ,并可杀伤这些细胞 ,半致死剂量为92ng/mL .
It is usually over-expressed on the surface of human breast cancer cells and other tumor cells that can be used as a target for tumor-directed therapy. Using genetic engineering techniques, the 6C6 single-chain antibody (ScFv6C6) is outside the Pseudomonas aeruginosa in which the deleted cell binds. Toxin A (PE40) is linked to form a 6C6 immunotoxin (ScFv6C6_PE40), in which ScFv6C6 is composed of the heavy chain variable region and the light chain variable region of a 6C6 monoclonal antibody that specifically recognizes the 28K protein. 6C6 immunotoxins The prokaryotic expression level was 3 3 %, approximately 55 mg/L broth. Purified using HisTrap (nickel ion chelate) column chromatography to give a purity of 3 32% of 6C6 immunotoxin, which can recognize MDA 2 3 human breast Tumor-associated antigens on the surface of cancer cells can kill these cells with a lethal dose of 92 ng/mL.