摘要
应用CelliGen细胞培养罐,培养抗人B血型杂交瘤细胞,获得了细胞密度超过10~7/ml,产物IgM 2g/L以上,特异性血凝效价达到2^(13)～2^(15)。培养条件50～80r/min、pH7.2、溶解氧(DO)30％～50％,高密度时维持DO2％～10％是必要的,4种气体混合流量为0.2LPM,高密度时要增加氧气供应,细胞密度达到1.6×10~6/ml时开始灌注,随细胞密度增加,灌注量亦增加,每天补充新培养液由800ml逐渐增加到3000ml。13B1杂交瘤细胞生长代谢与葡萄糖、乳酸有关,与氨代谢无关。
Hybridoma cell line 13Bl was cultured by CelliGen bioreactor. The product was more than 107/ml cells, IgM 2g/L, specific agglutinate titre 210 -215. The condition of culture was 37℃, RPM 50-80, pH 7.2 and Do 30-50%. The flow volume of mixed four kinds of air was 0.2 LPM. Perfusion was carried out when density of cells reached 1.6×106/ml. The quantity of perfusion was increased from 800 ml/day to 3000 ml/day, while the density of cells grew up. The relation between cell propagation and glucose lactatc was observed, but nothing to do with ammonia.
关键词
高密度培养
细胞培养罐
单克隆抗体
high density cell culture
antibody, monoclonal
human blood group B
bioreactor
