抗人A血型杂交瘤细胞高密度培养条件的研究

抗人 A 血型杂交瘤15B_4细胞在 CelliGen 细胞培养罐中进行连续灌注培养,细胞密度达1.6×10~7/ml。每天灌注新培养液量由1000ml 增到2300ml,第14天后又逐日减少至1000ml,死细胞数随着转速的增加和灌注量的减少而增多。血凝效价呈梯度上升,滴度达到2~(14),相当于腹水效价。培养6天 IgM 产量为0.2克/升/天,10～18天波动在1.45～1.8克/升/天。15B_4细胞代谢与葡萄糖、乳酸有关,与氨无关。CelliGen 自控效果较好,适用于杂交瘤细胞的高密度培养。搅拌速度在120～150r/min 对细胞产生的剪切力有明显地破坏作用。溶氧控制系统有待改进。

Study on Culture Condition of High Density of Hybridoma Cell Line Secreting Human Blood Group A Monoclonal Antibodies

Hybridoma cell line 15B_4 producing antibody against human blood group Awas cultured with cell culture perfusion in the bioreactor (Celligen).Cell density re-ached 1.6×10~7/ml and the amount of culture medium was increased from 1000ml to2300ml each day.The medium was reduced to.1000ml after 14th day,then the amountdead cells were increased with the decrease of perfusion quantity.Hemagglutinating ti-ter was greater to 2~(14) comparable with the antibody titer of ascites:The IgM yieldwas 1.6-1.86g/L.Metabolism of 15B_4 cells was related to concentration of glucoseand lactic acid,but not ammonia.The celligen was suitable for cell culture of hybridoma in high density,When sti-rring rate reached 150r/min,the cells were disrupted by shearing force.The dissolvedoxygen control system remains to be improved.