抗CD3 ScFv/抗PgP ScFv双功能抗体的高密度发酵与纯化  

High Cell Density Culture and Purification of AntiCD3 ScFvantiPgP ScFv Diabody

作　　者：王金宏[1] 刘娟妮[1] 纪庆[1] 高瀛岱[1] 熊冬生[1] 杨纯正[1] 

机构地区：[1]中国医学科学院中国协和医科大学血液学研究所实验血液学国家重点实验室,天津30002

出　　处：《药物生物技术》2008年第4期249-252,共4页Pharmaceutical Biotechnology

基　　金：天津市科技发展计划项目(No.05WFGPGX06900);国家自然科学基金资助项目(No.30400558)

摘　　要：在发酵罐上采用高密度发酵的方法对抗CD3ScFv/抗PgP ScFv双功能抗体工程菌进行了培养,其发酵结果是每升发酵液的湿菌菌重150g,OD550值达121;以免疫亲和层析的方法纯化抗CD3ScFv/抗PgPScFv双功能抗体,经计算抗体产量可达146.6mg/L;间接免疫荧光测定结果经流式细胞仪分析计算表明,抗CD3ScFv/抗PgP ScFv双功能抗体能与Jurkat细胞及K562/A02细胞特异性结合。Plasmid pAYZDCP was introduced to E. coli 16C9, in which it could be expressed at high level. By optimizing the ferment parameters, during high cell density culture the final cell density OD（550） reaches 121 （150 g wet cell weigh/L）. After the diabody was extracted from E. Coli and then purified by the immunoaffinity chromatography column. The yield of soluble antiCD3 ScFv/antiPgP ScFv diabody was 146. 6 mg/L. Its antigen binding activity, the diabody binding namely the activing of to Jurkat celIs（CD3^＋） and K562/A02 cells（PgP^＋） was examined by FACS. The affinities of the diabody were similar to the diabody which was obtained by shaking flask fermentation.

关 键 词：抗CD3 ScFv/抗PgP ScFv双功能抗体 高密度发酵 亲和层析 

分 类 号：Q936[生物学—微生物学]