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Abstract：
Peptide mapping with mass spectrometry (MS) is an important tool for protein characterization in the biopharmaceutical industry. Historically, peptide mapping monitors post-translational modifications (PTMs) of protein products and process intermediates during development. Multi-attribute monitoring (MAM) methods have been used previously in commercial release and stability testing panels to ensure control of selected critical quality attributes (CQAs). Our goal is to use MAM methods as part of an overall analytical testing strategy specifically focused on CQAs, while removing or replacing historical separation methods that do not effectively distinguish CQAs from non-CQAs due to co-elution. For example, in this study, we developed a strategy to replace a profile-based ion-exchange chromatography (IEC) method using a MAM method in combination with traditional purity methods to ensure control of charge variant CQAs for a commercial antibody (mAb) drug product (DP). To support this change in commercial testing strategy, the charge variant CQAs were identified and characterized during development by high-resolution LC-MS and LC-MS/MS. The charge variant CQAs included PTMs, high molecular weight species, and low molecular weight species. Thus, removal of the IEC method from the DP specification was achieved using a validated LC-MS MAM method on a QDa system to directly measure the charge variant PTM CQAs in combination with size exclusion chromatography (SE-HPLC) and capillary electrophoresis (CE-SDS) to measure the non-PTM charge variant CQAs. Bridging data between the MAM, IEC, and SE-HPLC methods were included in the commercial marketing application to justify removing IEC from the DP specification. We have also used this MAM method as a test for identity to reduce the number of QC assays. This strategy has received approvals from several health authorities.
摘要：
质谱（MS）的肽图是生物制药行业中蛋白质表征的重要工具。历史上,肽作图监测蛋白质产物的翻译后修饰（PTM）和开发过程中的中间体。多属性监测（MAM）方法先前已用于商业发布和稳定性测试面板，以确保控制选定的关键质量属性（CQA）。我们的目标是使用MAM方法作为专门针对CQA的整体分析测试策略的一部分，同时去除或替换由于共洗脱而不能有效区分CQAs和非CQAs的历史分离方法。例如,在这项研究中，我们开发了一种策略来替代基于谱的离子交换色谱(IEC)方法,该方法使用MAM方法与传统纯度方法相结合,以确保控制商业抗体(mAb)药物产品(DP)的电荷变体CQAs.为了支持商业测试策略的这一变化，在开发过程中通过高分辨率LC-MS和LC-MS鉴定和表征电荷变体CQAs。电荷变体CQA包括PTM，高分子量物种，和低分子量物种。因此，在QDa系统上使用经过验证的LC-MSMAM方法直接测量电荷变体PTMCQAs，并结合尺寸排阻色谱（SE-HPLC）和毛细管电泳（CE-SDS）测量非PTM电荷变体CQAs，从而从DP规范中去除IEC方法。在MAM之间桥接数据，IEC,和SE-HPLC方法包括在商业营销应用中，以证明从DP规范中删除IEC是合理的。我们还使用该MAM方法作为鉴定测试以减少QC测定的数量。这一战略已得到几个卫生当局的批准。
