Immobilized-enzyme reactors (IMERs) are flow-through devices containing enzymes that are physically confined or localized with retention of their catalytic activities. IMERs can be used repeatedly and continuously and have been applied for (bio)polymer degradation, proteomics, biomarker discovery, inhibitor screening, and detection. Online integration of IMERs with analytical instrumentation, such as high-performance liquid chromatography (HPLC) systems, reduces the time needed for multi-step workflows, reduces the need for sample handling, and enables automation. However, online integration can also be challenging, as reaching its full potential requires complex instrumental setups and experienced users. This review aims to provide an assessment of recent advances and challenges in online IMER-based (analytical) LC platforms, covering publications from 2014–2021. A critical discussion of challenges often encountered in IMER fabrication, sample preparation, integration into the analytical workflow, long-term usage, and of potential ways to overcome these is provided. Finally, the obstacles preventing the proliferation of IMERs as efficient tools for high-throughput pharmacological, industrial, and biological studies are discussed.

固定化酶反应器 (IMER) 是包含酶的流通装置，这些酶被物理限制或定位并保留其催化活性。IMER 可以重复和连续使用，并已应用于（生物）聚合物降解、蛋白质组学、生物标志物发现、抑制剂筛选和检测。IMER 与分析仪器（如高效液相色谱 (HPLC) 系统）的在线集成，减少了多步骤工作流程所需的时间，减少了对样品处理的需求，并实现了自动化。然而，在线集成也可能具有挑战性，因为充分发挥其潜力需要复杂的仪器设置和经验丰富的用户。本综述旨在评估基于在线 IMER（分析）LC 平台的最新进展和挑战，涵盖 2014-2021 年的出版物。对 IMER 制造、样品制备、分析工作流程集成、长期使用中经常遇到的挑战以及克服这些挑战的潜在方法进行了批判性讨论。最后，讨论了阻止 IMER 作为高通量药理学、工业和生物学研究的有效工具扩散的障碍。