【摘要】 转化了质粒PYZcpp3的 16C9大肠杆菌可高水平地分泌表达可溶性CD2 0F(ab′) 2 抗体 ;采用经优化的培养条件 ,在发酵罐上进行高密度培养OD550 值达 14 0 ;每升湿菌菌重 2 0 0g ;抗体的产量每升为 2 4 1mg ,其中F(ab′) 2 片段达 5 0 % ,F(ab′) 2 可以特异性的识别CD2 0 + 细胞并与CD2 0相结合 ;F(ab′) 2 对Raji细胞的IC50 值为 2 2 8μg mL ;而Fab′对Raji细胞的IC50 值为 4 5 9μg mL。更多还原

【Abstract】 The pYZcpp3 was introduced to E.coli 16C9,in which could be expressed at high level. By using optimize the ferment parameters ,during high density culture the final cell density OD-(550) reaches 140(200g wet cell weigh/L).After the CD20F(ab′)-2 antibody was extracted from E.coli and then purified by FPLC.The yield of soluble CD20F(ab′)-2 antibody was 241mg/L and 50%of it is bivalent. The antigen -binding activity of F(ab′)-2 was analysed by FACS using Raji cells as target cells,the result showed that F(ab′)-2 specifically binds to CD(20~+) cells.The activity of F(ab′)-2 inhibiting the growth of cell Raji in vitro was examined using MTT,which showed that the IC-(50) of CD20 F(ab′)-2 and Fab′ were (22.9μg/mL) and 42.8μg/mL respectively.更多还原