【摘要】 为探索电荷异构体的结构基础及其对生物学功能的影响,利用阳离子交换层析法分离制备了阿达木单抗的4个电荷异构体组分,通过液质联用进行结构表征,考察了生物学功能。结果表明,制备的电荷异构体具有较高的纯度、稳定性和相同的氨基酸序列;酸性变异体主要与聚集体、碎片、N-糖唾液酸化、脱酰胺水平升高和氧化比例降低有关;碱性变异体主要由聚集体、碎片、C-末端赖氨酸缺失不完全和氧化比例降低引起;电荷异质性未影响TNF-α结合活性和细胞死亡抑制活性。因此,电荷异构体不能被定义为杂质。更多还原

【Abstract】 In order to study structural information of charge variants and their effects on biological functions,adalimumab was separated by cation exchange chromatography to obtain four fractions of charge variants. The structures were characterized by LC-MS/MS, and the biological functions were investigated. The results show that the prepared charge variants had high purity, stability, and same amino acid sequence. The acidic variants were mainly related to aggregates, fragments, N-glycan sialylation, higher deamidation and lower oxidation,while the basic variants were caused by aggregates, fragments, incomplete removal of C-terminal lysine and decreased oxidation proportion. TNF-α binding affinities and cell death inhibitory activities were not affected by charge heterogeneity. Therefore, charge variants cannot be defined as impurities.更多还原