6C6鼠-人嵌合抗体的纯化及鉴定

采用基因工程技术 ,将小鼠 6C6单克隆抗体可变区基因与人抗体恒定区基因连接 ,构建了鼠 人 6C6嵌合抗体基因 ,并在CHO细胞中高效表达 .利用ProteinA亲和层析柱从细胞培养上清中分离纯化 6C6嵌合抗体 ,得到电泳纯度大于 98%的 6C6嵌合抗体 ,其重链 (5 5kD)和轻链 (2 4kD)符合IgG相对分子质量的理论值 .Western印迹、细胞免疫荧光和免疫组织化学实验结果均呈阳性 .表明6C6嵌合抗体可识别人乳腺癌细胞表面上的肿瘤相关抗原 ,保持了 6C6单克隆抗体的特性 ,为后续的研究工作奠定了基础

Purification and Characterization of 6C6 Mouse-human Chimeric-antibody

A mouse-human 6C6 chimeric antibody gene, which is a linkage of variable region gene of mouse 6C6 monoclone antibody and constant region gene of human antibody, has been cloned and effectively expressed in CHO cells. The 6C6 chimeric antibody with a purity of more than 98% identified by electrophoresis was obtained from the cell culture supernatants by Protein A affinity chromatography. Both the heavy chain (55 kD) and light chain (24 kD) of the antibody were in accordance with the theoretical molecular weight of IgG. The positive results of Western blotting, cell immunofluorescence and immunohistochemistry tests indicated that the 6C6 chimeric antibody could recognize tumor-associated antigen on human breast cancer cell surface and thus had the characteristics of mouse 6C6 monoclone antibody.