Abstract：
Monoclonal antibodies (mAbs) represent the largest class of therapeutic protein drug products. mAb glycosylation produces a heterogeneous, analytically challenging distribution of glycoforms that typically should be adequately characterized because glycosylation-based product quality attributes (PQAs) can impact product quality, immunogenicity, and efficacy. In this study, two products were compared using a panel of analytical methods. Two high-resolution mass spectrometry (HRMS) workflows were used to analyze N-glycans, while nuclear magnetic resonance (NMR) was used to generate monosaccharide fingerprints. These state-of-the-art techniques were compared to conventional analysis using hydrophilic interaction chromatography (HILIC) coupled with fluorescence detection (FLD). The advantages and disadvantages of each method are discussed along with a comparison of the identified glycan distributions. The results demonstrated agreement across all methods for major glycoforms, demonstrating how confidence in glycan characterization is increased by combining orthogonal analytical methodologies. The full panel of methods used represents a diverse toolbox that can be selected from based on the needs for a specific product or analysis.
摘要：
单克隆抗体(mAb)代表最大类别的治疗性蛋白质药物产品。mAb糖基化产生异质，由于基于糖基化的产品质量属性（PQA）可能影响产品质量，因此通常应充分表征的糖型分析具有挑战性的分布，免疫原性,和功效。在这项研究中,使用一组分析方法比较了两种产品。两个高分辨率质谱（HRMS）工作流程用于分析N-聚糖，而核磁共振(NMR)用于产生单糖指纹图谱。将这些现有技术与使用结合有荧光检测(FLD)的亲水相互作用色谱(HILIC)的常规分析进行比较。讨论了每种方法的优缺点，并比较了已鉴定的聚糖分布。结果表明，所有方法对主要糖型的一致性，证明如何通过结合正交分析方法提高聚糖表征的信心。所使用的方法的完整面板代表了一个不同的工具箱，可以根据特定产品或分析的需求进行选择。
