运用全因子试验方法优化单抗生物类似药生产过程中的糖基化分布

Optimization of Glycosylation Distribution in the Production of Monoclonal Antibody Bio-Medicine by Full Factorial Design

旨在调控单抗生物类似药的糖基化分布,使其与原研药一致。通过全因子设计试验考察不同浓度的半乳糖、尿苷、氯化锰和蛋白水解物对单抗糖基化修饰的影响。结果表明,半乳糖、尿苷及氯化锰均能提高单抗半乳糖基化,且对细胞生长和单抗产量无明显影响。此外尿苷和氯化锰还能降低五聚高甘露糖型(Man5),但过量的尿苷会导致Man5增高。而蛋白水解物除了能降低Man5,提高单抗岩藻糖基化外,对细胞生长也有促进作用。选取模型预测的最优条件进行反应器培养验证,最终的单抗糖基化分布符合预期,成功建立CHO细胞单抗糖基化调控的生产策略。

This work aims to regulate the glycosylation distribution of monoclonal antibody bio-medicine,and thus which allows them to be in accord with original medicine.Full factorial design was applied to investigate the effects of 4 additives of galactose,uridine,manganese chloride and hydrolysate on the glycosylation of monoclonal antibody(mAb).The results revealed that combined supplements of galactose,uridine and manganese chloride increased galactosylation of mAb;moreover,there was no significant impact on cell growth and antibody production.Furthermore,uridine and manganese chloride inhibited Man5 formation while excessive uridine led to a high Man5 level.Besides,protolysate not only decreased Man5 level and increased fucosylation,also enhanced cell growth.In summary,a desired target glycosylation profile was achieved after the validation was conducted through bioreactor cultivation under the optimized condition predicted by a model,thus production strategy of regulating the glycosylation of m Ab during CHO cell culture process is successfully established.