【摘要】 通过逐步降低血清浓度,HPLC氨基酸分析及正交实验筛选研制了HAb18杂交瘤细胞的无血清培养基。对在该无血清培养条件下的细胞进行了计数,对培养上清液进行了葡萄糖、谷氨酰胺、乳酸和氨浓度以及抗体分泌量和抗原结合活性测定,并对动力学参数进行了分析,结果表明HAb18细胞在无血清培养条件下达到的最大细胞密度和抗体分泌量分别为0.91×106个/ml和43.8mg/L;细胞比生长速率较在有血清条件下稍有下降,而抗体合成速率提高(0.0207/h比0.0218/h,0.387pg/cell/h比0.218pg/cell/h,P<0.01)。无血清培养时葡萄糖和谷氨酰胺消耗无明显变化,但乳酸浓度降低,氨浓度升高;此外,分泌抗体的抗原结合活性增加。研究无血清培养条件下的HAb18细胞生长代谢和抗体分泌特征可为建立HAb18无血清悬浮流加培养工艺打下基础。更多还原

【Abstract】 A serum free medium for hybridoma HAb18 was obtained through decreasing the serum concentration gradually, analyzing amino acids exhaustion with HPLC and screening serum supplement through orthogonal experiment. Cell grown in serum free medium was subjected to cell counting. Supernatant from the spent medium were pooled to determine the concentration of glucose, glutamine, lactate and ammonia. Moreover, not only antibody yield and antigen binding activity were detected, but dynamic parameters were analyzed. Our results showed that the maximum cell density and antibody production in serum free medium were 0.91×106 cells/ ml and 43.8 mg/L, respectively. Although the specific cell growth rate reduced slightly, antibody productivity increased (0.0207/h vs. 0.0218/h, 0.387 vs. 0.218 pg/cell/h, P<0.01). Glucose and glutamine exhaustions in serum free medium were similar to that in serum containing medium; while lactate production cut down and ammonia production enhanced. Likewise, antigen-binding activity increased. Taken together, studies on characteristics of HAb18 cell growth, metabolism and antibody synthesis in serum free medium would lay a foundation for HAb18 serum free suspension fed-batch technology.更多还原