Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis
Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis
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DOI：
10.1021/acs.analchem.6b04726
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发表时间：
2017-02-07
影响因子：
7.4
通讯作者：
Guillarme, Davy
中科院分区：
文献类型：
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作者：
D'Atri, Valentina;Fekete, Szabolcs;Guillarme, Davy
生物类似药mAb的开发和批准过程取决于其与原研药的可比性。因此，需要进行分析比较，以评估结构特征和翻译后修饰（PTM），从而最大限度地降低生物类似药和原研药之间具有临床意义差异的风险。mAb的糖基化模式被认为是一个重要的关键质量属性（CQA），已经提出了几种分析方法来促进糖基化特征的表征和监测，尽管主要是在聚糖和糖肽水平的分析。在本研究中，我们通过使用中间方法比较原研药和生物仿制药mAb（类克/Remsina/Inflectra、赫赛汀/曲妥珠单抗B和爱必妥/西妥昔单抗B），证明了亲水相互作用色谱（HILIC）与质谱（MS）联用在蛋白质水平上定性分析糖基化模式的实用性。我们证明了HILIC解决蛋白质生物药物的亲水性变体的能力，在分析的中间水平，它的互补性，反相液相色谱法，和它的连字符到MS。HILIC功能结合到MS作出了一个强大的分析工具，用于比较原始和生物类似单克隆抗体，最终可以应用于常规分析的质量控制。
The development and approval processes of biosimilar mAbs depend on their comparability to originators. Therefore, analytical comparisons are required to assess structural features and post translational modifications (PTM) and thereby minimize the risk of being clinically meaningful differences between biosimilar and originator drug products. The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control.