| 摘要: |
| 蛋白多肽是一类具有生物活性的物质，对维持机体各项功能的稳定发挥着至关重要的作用，快速、准确地检测其水平有助于疾病的诊断、药物治疗效果的监测及药物的研发等，在临床医学、生物学及药学等领域具有重要意义。传统的蛋白多肽检测方法（如蛋白质印迹法和ELISA等）存在灵敏度低、难以同时测定多个待测物等问题。液相色谱-质谱法（LC-MS）具有专属、灵敏、高通量等优势，但蛋白多肽大分子的低离子化效率及强基质效应等限制了其直接检测蛋白多肽的可行性，基于LC-MS的信号转化与放大策略应运而生。本文针对近年来基于LC-MS分析生物体内蛋白多肽的样品前处理方法和信号转化与放大策略的研究进展进行归纳总结，为基于LC-MS开发复杂样品中低丰度蛋白多肽的专属高敏检测方法提供参考。 |
| 关键词: 蛋白多肽 液相色谱-质谱法 样品前处理 信号转化与放大 |
| DOI：10.16781/j.CN31-2187/R.20230643 |
| 投稿时间：2023-11-20修订日期：2024-01-18 |
| 基金项目:国家自然科学基金(82104124)，上海市科学技术委员会“科技创新行动计划”扬帆计划（20YF1458500），上海市“科技创新行动计划”生物医药科技支撑专项（22S21904000）. |
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| Liquid chromatography-mass spectrometry analysis for low-abundance protein polypeptides in vivo: research progress |
| GENG Shuwen1,2,YANG Wen1*,MAO Junqin2,ZHOU Tingting1 |
(1. Department of Pharmaceutical Analysis, School of Pharmacy, Naval Medical University (Second Military Medical University), Shanghai 200433, China;
2. Department of Clinical Pharmacy, Jiading Branch of Shanghai General Hospital (Jiangqiao Hospital of Jiading District), Shanghai 201803, China
*Corresponding author) |
| Abstract: |
| Protein polypeptides are a class of bioactive substances that play a crucial role in maintaining the stability of various functions of the organism. Rapid and accurate detection of their levels could help in the diagnosis of diseases, the monitoring of drug therapy and the research and development of medicines, which is of great significance in the fields of clinical medicine, biology and pharmacy. Conventional protein polypeptides detection methods, such as Western blotting and enzyme-linked immunosorbent assay, still have problems like low sensitivity or difficulty in determining more than 1 analyte simultaneously. Liquid chromatography-mass spectrometry (LC-MS) has the advantages of specificity, sensitivity and high throughput. However, low ionization efficiency of macromolecules and strong biological matrix effects limit the feasibility of direct detection of protein polypeptides by LC-MS, which has led to the development of signal conversion and amplification strategies based on LC-MS technology. In this review, we summarized the research progress of sample pretreatment methods and signal conversion and amplification strategies for quantification of protein polypeptides in vivo based on LC-MS in recent years, providing a reference for developing specific high-sensitivity detection methods for low-abundance protein polypeptides in complex samples based on LC-MS technology. |
| Key words: protein polypeptides liquid chromatography-mass spectrometry sample pretreatment signal conversion and amplification |
