抗P185工程抗体表达与纯化研究  

The Research on Expression and Purification of a Chimeric Anti-p185 Antibody

作　　者：乔婧娟[1] 刘海波[1] 朱娟娟[1] 程联胜[1] 刘兢[1] 

机构地区：[1]中国科学技术大学生命科学学院,合肥230027

出　　处：《中国生物工程杂志》2006年第9期32-37,共6页China Biotechnology

基　　金：国家"863"计划资助项目(2004AAZ15260);国家自然科学基金资助项目(30570362)

摘　　要：人类的原癌基因Her-2编码一个185kDa的跨膜受体酪氨酸激酶,也称为p185,是肿瘤相关蛋白,在多种癌细胞,特别在乳腺癌中有高表达。构建了抗p185/HER-2的二价的嵌合抗体(scFv-Fc),并证明其保留了亲本抗体的特异结合活性,进而建立了高表达的细胞株。对该嵌合抗体进行滚瓶大规模培养,并对培养产物的纯化条件进行了摸索,建立了亲和层析和阳离子交换层析两步法。最终产物的纯度在95%以上,回收率为63%。摸索了嵌合抗体的冻干保存条件,抗体可以保持其稳定性和结合活性一年以上。嵌合抗体纯化工艺和保存条件的确定,为进一步的临床研究奠定了基础。The Her-2 proto-oncogene encodes a 185kDa transmenbrane glycoprotein p185 which has intrinsic tyrosine kinase activity. It is overexpressed in several malignant human tumors like breast cancer. A chimeric antibody by assembling a single-chain Fv antibody and a human IgG1 Fc fragment was constructed. This chimeric antibody reacts with tumor surface antigen p185c-erbB-2 specifically. In order to put the antibody into clinical application, two steps purification method was used to attain the antibody＇ s purity more than 95%. Both the lyophilized pharmaceutical formulations of the antibody were found. The formulations can keep the stability and activity of the antibody for at least one year. These results were the foundation of the chimeric antibody for cancer therapy.

关 键 词：嵌合抗体 亲和纯化 离子交换纯化 稳定性 

分 类 号：Q819[生物学—生物工程]