在中空纤维细胞培养系统内用无血清培养基生产单克隆抗体  

Production of Monoclonal Antibody in Hollow Fiber Culture System with Serum-free Medium

作　　者：周维松[1] 蔡少华[1] 

机构地区：[1]中国农业科学院生物技术研究中心

出　　处：《生物工程学报》1992年第1期70-76,共7页Chinese Journal of Biotechnology

基　　金：农业部资助项目

摘　　要：用1640SFM无血清培养基在VF-2中空纤维细胞培养系统内培养7E8杂交瘤细胞,最大活细胞密度为2.34×10~6/ml,该活细胞密度分别是转瓶培养和静态瓶培养结果的3.7倍和2.2倍。培养42天共收获7E8细胞培养液10000ml,其单克隆抗体(简称单抗)的ELISA效价为1:20000左右,该效价是转瓶培养和静态瓶培养结果的25倍。1000ml培养液经50%饱和硫酸铵盐析和Sephadex G-200柱层析提纯,获纯化单抗IgG 51.82mg。该系统平均每天生产单抗12.3mg。研究结果表明:在中空纤维培养系统内用无血清培养基培养杂交瘤细胞的方法可望用于体外大规模生产单抗。Hybridoma 7E8 cells, which produced IgG_(2a) aganist potato virus X, were cultured in VF-2 hollow fiber cell culture system with 1640 SFM serum-free medium. The maximam viable cell density was 2.34×10~6 cells/ ml that was 3.7 times and 2.2 times of the maximam viable cell density achieved in spinner flask culture system and in static flask culture system, respectively. 10000 ml cell cultural supernatant had been collected in 42 days cultural period. The ELISA titer of monoclonal antibody (McAb)in the supernatant was about 1:20000 that was 25 times of the titers attained in spinner flask culture system and in static flask culture system. From 1000ml cell cultural supernatant, an amount of 51.2mg 7E8 McAb IgG had been purified by 50% saturated ammonium sulfate precipitation and Sephadex G-200 chromatography. The average yield of McAb in the hollow fiber cell culture system was 12.3mg/d. The results of the study suggest that it is hopeful fol bybridoma cell culture in the hollow fiber cell culture system with serum-free medium for McAb production on larger-scale.

关 键 词：单克隆抗体 中空纤维系统 杂交瘤 

分 类 号：S188[农业科学—农业基础科学]