杂交瘤细胞在CelliGen培养器中大量连续培养

A Perfusion System for High Productivity of Monoclonal Antibody by Hybridoma Cells in A CelliGen Bioreactor

鼠-鼠杂交瘤细胞DA4-4(ATCC,HB57)分泌抗人IgM的单克隆抗体IgGl,该细胞被培养在1.5L CelliGen-中空纤维柱连续灌注系统中,CelliGen细胞培养器显示很低的机械剪切力,提高搅拌速度到150rpm增加了氧的传递,但不对杂交瘤细胞造成损害。在连续培养时,DA4-4细胞密度可达20×10~6/ml以上,单克隆抗体浓度高达4.75mg/ml,该培养系统每日可收获抗体1.0g左右。

DA4-4 (ATCC HB57)is a shear sensitive mouse-mouse hybridoma cell, producing monoclonal antibody IgG1 against human IgM. It was grown successfully in a perfusion propagation system consisting of a 1.5L CelliGen stirred bioreactor and Hollow fiber cartridge. CelliGen system had low shear forces and cells can grow well at high agitation of 150 rpm. The culture was maintained for 40 days and cell number reached 32×10~6/ml. Maximal monoclonal antibody concetration was 4.75mg/ml. This system produced about 1.0g of antibody per day at its pesks.