采用混合培养基模式将杂交瘤细胞进行减血清悬浮驯化培养，通过优化提高单克隆抗体的表达量，并进行抗体的分离纯化。在杂交瘤细胞悬浮培养优化过程中，最终确定的血清浓度为2％胎牛血清，培养基配比为RPM1640∶SFM4CHO∶OptiCHO（4.5∶4.5∶1），接种密度为3×105 cells／mL，采用批次补料流加培养，每2 d流加1次，每次流加量为初始体积的3％，抗体的表达量从原始的84 mg／L提高到168 mg／L。将其进一步在5 L搅拌式生物反应器中进行培养，离心后上清经protein A亲和层析纯化，确定抗体的产量为240 mg／L。%In this study , hybridoma cell was adapted to the mixed medium containing less fetal bovine serum through suspension do -mestication, and the antibody concentration increased largely .The components of mixed medium were RPM 1640 ∶SFM4CHO∶Opti-CHO(4.5∶4.5∶1) with 2%FBS and the fed-batch fermentation process was carried out , in which the initial inoculation concentration was 3 ×105 cells/mL, and 3%(initial volume) chemical-defined feed medium was added every other day during cultivation .The anti-body concentration increased to 168 mg/L from 84 mg/L.The hybridoma cells were further cultured in a 5 L stirred-bioreactor , and the products were purified by protein A affinity chromatography and the purified antibody concentration was 240 mg/L.
展开▼
