Therapeutic monoclonal antibodies (mAbs) are highly heterogeneous as a result of posttranslational modifications (PTMs) during bioprocessing and storage. The modifications that impact mAb product quality are regarded as critical quality attributes and require monitoring. The conventional LC–mass spectrometer (MS) method used for product quality monitoring may require protein A purification prior to analysis. In this paper, we present a high-throughput microchip electrophoresis (<4 min) in-line with MS (MCE–MS) that enables baseline separation and characterization of Fc, Fd′, and light chain (LC) domains of IdeS-treated mAb sample directly from bioreactor. The NISTmAb was used to optimize the MCE separation and to assess its capability of multiple attribute monitoring. The MCE–MS can uniquely separate and characterize deamidated species at domain level compared to LC–MS method. Two case studies were followed to demonstrate the method capability of monitoring product quality of mAb samples from stability studies or directly from bioreactors.

中文翻译：

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微芯片电泳分离与质谱联用 (MCE–MS) 用于快速监测单克隆抗体的多种质量属性

由于生物加工和储存过程中的翻译后修饰 (PTM)，治疗性单克隆抗体 (mAb) 具有高度异质性。影响 mAb 产品质量的修饰被视为关键质量属性，需要进行监控。用于产品质量监测的常规 LC-质谱仪 (MS) 方法可能需要在分析前对蛋白 A 进行纯化。在本文中，我们提出了一种与 MS (MCE–MS) 联机的高通量微芯片电泳（<4 分钟），可实现 IdeS 处理的 Fc、Fd' 和轻链 (LC) 结构域的基线分离和表征mAb 样品直接来自生物反应器。NISTmAb 用于优化 MCE 分离并评估其多属性监测能力。与 LC-MS 方法相比，MCE-MS 可以在域水平上独特地分离和表征脱酰胺物质。随后进行了两个案例研究，以证明方法能够监测来自稳定性研究或直接来自生物反应器的 mAb 样品的产品质量。