Recent advances in analytical techniques provide the opportunity to quantify even low-abundance glycopeptides derived from complex biological mixtures, allowing for the identification of glycosylation differences between healthy samples and those derived from disease states. Herein, we discuss the sample preparation procedures and the mass spectrometry (MS) strategies that have facilitated glycopeptide quantification, as well as the standards used for glycopeptide quantification. For sample preparation, various glycopeptide enrichment methods are summarized including the columns used for glycopeptide separation in liquid chromatography separation. For MS analysis strategies, MS1 level-based quantification and MS2 level-based quantification are described, either with or without labeling, where we have covered isotope labeling, TMT/iTRAQ labeling, data dependent acquisition, data independent acquisition, multiple reaction monitoring, and parallel reaction monitoring. The strengths and weaknesses of these methods are compared, particularly those associated with the figures of merit that are important for clinical biomarker studies and the pathological and functional studies of glycoproteins in various diseases. Possible future developments for glycopeptide quantification are discussed.

中文翻译：

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通过液体分离和质谱法定量糖肽的方法

分析技术的最新进展提供了量化来自复杂生物混合物的低丰度糖肽的机会，从而可以识别健康样品和来自疾病状态的样品之间的糖基化差异。在此，我们讨论了促进糖肽定量的样品制备程序和质谱 (MS) 策略，以及用于糖肽定量的标准。对于样品制备，总结了各种糖肽富集方法，包括液相色谱分离中用于糖肽分离的色谱柱。对于 MS 分析策略，描述了基于 MS1 水平的量化和基于 MS2 水平的量化，有或没有标记，其中我们涵盖了同位素标记、TMT/iTRAQ 标记、数据依赖采集、数据独立采集、多反应监测和平行反应监测。比较了这些方法的优点和缺点，特别是那些与品质因数相关的方法，这些品质因数对临床生物标志物研究以及各种疾病中糖蛋白的病理学和功能研究很重要。讨论了糖肽定量的未来可能发展。