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利用鼠鼠杂交瘤2F7细胞(分泌IgG2a单抗)研究了Pluronic F-68、甲基纤维素、羧甲基纤维素及聚醚多元醇与杂交瘤细胞生物相容性及添加限制浓度;研究了添加浓度对葡萄糖的利用及氨的生成影响;在高速搅拌、高剪切力下考查添加剂的保护效果。结果表明,0.05—0.10％(W/V)Pluronic F-68、0.10—0.20％(W/V)甲基纤维素能较好地保护杂交瘤细胞;高浓度Pluronic F-68增加了葡萄糖的比消耗速率及氨的比生成速率;高浓度甲基纤维素增加了氨比生成速率;羧甲基纤维素添加浓度低于0.1％不影响细胞生长,也无保护作用,羧甲基纤维素不影响细胞对葡萄糖的比消耗速率,但增加了氨比生成速率;聚醚多元醇分解细胞。在1.5升GelliGen生物反应器中,培养基添加0.10％Pluronic F-68、搅拌转速70r/min下细胞正常生长。
The biocompatibility and limiting concentration of Pluronic F-68, methylcellulose, carboxymethylcellulose and polyether polyols and hybridoma cells were studied using mouse and mouse hybridoma 2F7 cells (secreted IgG2a monoclonal antibody); The effect of adding concentration on the utilization of glucose and the formation of ammonia; the protective effect of additives was examined under high-speed stirring and high shear force. The results showed that 0.05-0.10% (W/V) Pluronic F-68, 0.10-0.20% (W/V) methylcellulose could better protect hybridoma cells; high concentrations of Pluronic F-68 increased glucose ratio Consumption rate and ammonia ratio generation rate; high concentration of methyl cellulose increased the rate of ammonia ratio generation; carboxymethyl cellulose added concentration of less than 0.1% did not affect cell growth, nor protective effect, carboxymethyl cellulose did not affect The rate of specific glucose consumption by the cells, but increased the rate of ammonia production; polyether polyols break down cells. In a 1.5 liter GelliGen bioreactor, cells were grown normally with 0.10% Pluronic F-68 and agitation at 70 r/min.