应用自动化和高通量 MALDI 质谱法进行多肽代谢稳定性筛选,Journal of the American Society for Mass Spectrometry

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应用自动化和高通量 MALDI 质谱法进行多肽代谢稳定性筛选
Journal of the American Society for Mass Spectrometry ( IF 3.1 ) Pub Date : 2023-05-17 , DOI: 10.1021/jasms.3c00091
Huifang Yao ₁ , Bingming Chen ₂ , Paul Harradine ₂ , Bahanu Habulihaz ₂ , David G. McLaren ₁ , Mark T. Cancilla ₂

Affiliation

肽疗法的发现代表了药物研究中一个快速增长的部分。在早期发现过程中，需要快速筛选大量候选肽在相关生物基质中的代谢稳定性。在大多数情况下，肽稳定性测定是使用 LC–MS/MS 进行量化的，这可能需要数小时来分析 384 个样品并产生数升的溶剂废物。在此，我们介绍了一种基于基质辅助激光解吸/电离 (MALDI) 质谱 (MS) 的肽稳定性评估高通量筛选 (HTS) 平台。样品制备已实现全自动化，人工干预最少。评估了平台的检测限、线性和重现性，并确定了许多候选肽的代谢稳定性。基于 MALDI-MS 的 HTS 工作流程能够在不到 1 小时内分析 384 个样品，同时仅使用 115 μL 的总溶剂。尽管此过程允许非常快速地评估肽稳定性，但鉴于 MALDI 过程的性质，值得注意的是观察到点对点变化和电离偏差。因此，可能仍然需要 LC-MS/MS 进行可靠的定量测量和/或当使用 MALDI 时某些肽的电离效率不足。

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Applying Automation and High-Throughput MALDI Mass Spectrometry for Peptide Metabolic Stability Screening

The discovery of peptide therapeutics represents a fast-growing segment of pharmaceutical research. During the early discovery process, a large number of peptide candidates needs to be rapidly screened for metabolic stability in relevant biological matrices. In most cases, peptide stability assays are quantified using LC–MS/MS, which may take hours to analyze 384 samples and generates liters of solvent waste. Herein, we introduce a high-throughput screening (HTS) platform for peptide stability assessment founded on Matrix Assisted Laser Desorption/Ionization (MALDI) mass spectrometry (MS). Full automation has been implemented for sample preparation with minimal manual intervention. The limit of detection, linearity, and reproducibility of the platform were evaluated, and metabolic stabilities have been determined for a number of peptide candidates. The MALDI-MS-based HTS workflow is able to analyze 384 samples in less than 1 h while only using 115 μL of total solvent. Although this process allows for very rapid assessment of peptide stability, given the nature of the MALDI process, it is noteworthy that spot-to-spot variations and ionization bias are observed. Therefore, LC–MS/MS may still be needed for confident, quantitative measurements and/or when the ionization efficiency of certain peptides is inadequate using MALDI.

更新日期：2023-05-17

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