Center for Intelligent Drug Delivery and Sensing Using Microcontainers and Nanomechanics (IDUN), Department of Health Technology, Technical University of Denmark, Kgs. Lyngby 2800, Denmark.
BioInnovation Institute Foundation, Copenhagen N 2200, Denmark.
ACS Sens. 2022 Aug 26;7(8):2358-2369. doi: 10.1021/acssensors.2c01022. Epub 2022 Jul 18.
Therapeutic drug monitoring (TDM) is an essential clinical practice for optimizing drug dosing, thereby preventing adverse effects of drugs with a narrow therapeutic window, slow clearance, or high interperson pharmacokinetic variability. Monitoring methotrexate (MTX) during high-dose MTX (HD-MTX) therapy is necessary to avoid potentially fatal side effects caused by delayed elimination. Despite the efficacy of HD-MTX treatment, its clinical application in resource-limited settings is constrained due to the relatively high cost and time of analysis with conventional analysis methods. In this work, we developed (i) an electrochemically assisted surface-enhanced Raman spectroscopy (SERS) method for detecting MTX in human serum at a clinically relevant concentration range and (ii) a benchtop, Raman detection system with an integrated potentiostat, software, and data analysis unit that enables mapping of small areas of SERS substrates and quantitative SERS-based analysis. In the assay, by promoting electrostatic attraction between gold-coated nanopillar SERS substrates and MTX molecules in aqueous samples, a detection limit of 0.13 μM with a linear range of 0.43-2 μM was achieved in PBS. The implemented sample cleanup through gel filtration proved to be highly effective, resulting in a similar detection limit (0.55 μM) and linear range (1.81-5 μM) for both PBS and serum. The developed and optimized assay could also be used on the in-house built, Raman device. We showed that MTX detection can be carried out in less than 30 min with the Raman device, paving the way toward the TDM of MTX at the point-of-need and in resource-limited environments.
治疗药物监测（TDM）是优化药物剂量的重要临床实践，可预防治疗窗较窄、清除率低或个体间药代动力学变异性较大的药物的不良反应。在大剂量甲氨蝶呤（HD-MTX）治疗期间监测甲氨蝶呤（MTX），以避免因延迟消除而导致潜在的致命副作用。尽管 HD-MTX 治疗有效，但由于常规分析方法的分析成本和时间相对较高，其在资源有限的环境中的临床应用受到限制。在这项工作中，我们开发了（i）一种电化学辅助表面增强拉曼光谱（SERS）方法，用于检测人血清中临床相关浓度范围内的 MTX，以及（ii）一种带有集成电化学工作站、软件和数据分析单元的台式拉曼检测系统，可实现 SERS 基底的小面积映射和基于 SERS 的定量分析。在该测定中，通过促进金包被纳米柱 SERS 基底和水溶液样品中 MTX 分子之间的静电吸引，在 PBS 中实现了 0.13 μM 的检测限和 0.43-2 μM 的线性范围。通过凝胶过滤实施的样品净化被证明非常有效，导致 PBS 和血清中的检测限（0.55 μM）和线性范围（1.81-5 μM）相似。开发和优化的测定法也可以在内部构建的拉曼设备上使用。我们表明，使用拉曼设备可以在不到 30 分钟内完成 MTX 检测，为在即时和资源有限的环境中进行 MTX 的 TDM 铺平了道路。
