This review provides an overview of the latest advancements and applications in multi‐dimensional liquid chromatography coupled with mass spectrometry (mD‐LC‐MS), covering aspects such as inter‐laboratory studies, digestion strategy, trapping column, and multi‐level analysis. The shift from an offline to an online workflow reduces sample processing artifacts, analytical variability, analysis time, and the labor required for data acquisition. Over the past few years, this technique has demonstrated sufficient maturity for application across a diverse range of complex products. Moreover, there is potential for this strategy to evolve into an integrated process analytical technology tool for the real‐time monitoring of monoclonal antibody quality. This review also identifies emerging trends, including its application to new modalities, the possibility of evaluating biological activity within the mD‐LC set‐up, and the consideration of multi‐dimensional capillary electrophoresis as an alternative to mD‐LC. As mD‐LC‐MS continues to evolve and integrate emerging trends, it holds the potential to shape the next generation of analytical tools, offering exciting possibilities for enhanced characterization and monitoring of complex biopharmaceutical products.

中文翻译：

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多维技术——生物治疗表征的最新进展和应用

本综述概述了多维液相色谱与质谱联用 (mD-LC-MS) 的最新进展和应用，涵盖实验室间研究、消化策略、捕获柱和多级分析等方面。从离线到在线工作流程的转变减少了样品处理工件、分析变异性、分析时间和数据采集所需的劳动力。在过去的几年中，该技术已经表现出足够的成熟度，可以应用于各种复杂的产品。此外，该策略有可能发展成为实时监测单克隆抗体质量的集成过程分析技术工具。本综述还确定了新兴趋势，包括其在新模式中的应用、评估 mD-LC 装置内生物活性的可能性，以及考虑多维毛细管电泳作为 mD-LC 的替代品。随着 mD-LC-MS 不断发展并整合新兴趋势，它具有塑造下一代分析工具的潜力，为增强复杂生物制药产品的表征和监测提供令人兴奋的可能性。