地诺单抗生物类似药的N-糖型优化

N-glycosylation optimization for biosimilar denosumab

目的:使用糖型调节试剂优化地诺单抗(denosumab)生物类似药的N-糖基化修饰,使其主要糖型G0F和G1F的比例与地诺单抗高度类似。方法:利用单因素和DOE试验方法,考察不同浓度氯化锰、半乳糖及尿苷对地诺单抗生物类似药糖基化修饰的影响。结果:经过单因素试验、DOE分析及200 L细胞培养工艺放大,确定糖型调节剂氯化锰和半乳糖的最佳用量分别是150μmol/L和35.56 mmol/L。结论:使用糖型调节剂氯化锰和半乳糖能有效调节地诺单抗生物类似药的糖基化修饰比例,可为其他单抗生物类似药生产工艺的优化提供借鉴。

Objective:To optimize the N-glycosylation of denosumab biosimilar using some glycosylation reagents so as to make the proportion of G0 F and G1 F be highly similar to denosumab.Methods:The effects of the different concentrations of manganese chloride、galactose and uridine on the glycosylation of denosumab biosimilar were studied by single factor and DOE assays.Results:A preferred combination including manganese chloride 150μmol/L,galactose 35.56 mmol/L was obtained by single factor test,DOE analysis and 200 L cell cultivation.Conclusion:The use of manganese chloride and galactose can effectively regulate the glycosylation modification ratio of denosumab biosimilar,which can provide a reference for the optimization of production process of the other mono-cloning antibody biosimilars.