Glycans introduce complexity to the proteins to which they are attached. These modifications vary during the progression of many diseases; thus, they serve as potential biomarkers for disease diagnosis and prognosis. The immense structural diversity of glycans makes glycosylation analysis and quantitation difficult. Fortunately, recent advances in analytical techniques provide the opportunity to quantify even low-abundant glycopeptides and glycans derived from complex biological mixtures, allowing for the identification of glycosylation differences between healthy samples and those derived from disease states. Understanding the strengths and weaknesses of different quantitative glycomics analysis methods is important for selecting the best strategy to analyze glycosylation changes in any given set of clinical samples. To provide guidance towards selecting the proper approach, we discuss four widely used quantitative glycomics analysis platforms, including fluorescence-based analysis of released N-linked glycans and three different varieties of MS-based analysis: liquid chromatography (LC)-mass spectrometry (MS) analysis of glycopeptides, matrix-assisted laser desorption ionization-time of flight MS, and LC-ESI-MS analysis of released N-linked glycans. These methods' strengths and weaknesses are compared, particularly associated with the figures of merit that are important for clinical biomarker studies, including: the initial sample requirements, the methods' throughput, sample preparation time, the number of species identified, the methods' utility for isomer separation and structural characterization, method-related challenges associated with quantitation, repeatability, the expertise required, and the cost for each analysis. This review, therefore, provides unique guidance to researchers who endeavor to undertake a clinical glycomics analysis by offering insights on the available analysis technologies.

中文翻译：

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定量临床糖组学策略：选择最佳分析方法的指南

聚糖给它们所附着的蛋白质带来了复杂性。这些修饰在许多疾病的进展过程中会发生变化；因此，它们可作为疾病诊断和预后的潜在生物标志物。聚糖的巨大结构多样性使得糖基化分析和定量变得困难。幸运的是，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和聚糖的机会，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。了解不同定量糖组学分析方法的优缺点对于选择最佳策略来分析任何给定的一组临床样本中的糖基化变化非常重要。N-连接聚糖和三种不同类型的基于 MS 的分析：糖肽的液相色谱 (LC)-质谱 (MS) 分析、基质辅助激光解吸电离-飞行时间 MS 和释放的 LC-ESI-MS 分析否-连接的聚糖。比较了这些方法的优点和缺点，特别是与对临床生物标志物研究很重要的品质因数相关的方法，包括：初始样品要求、方法的通量、样品制备时间、鉴定的物种数量、方法的效用对于异构体分离和结构表征、与定量相关的方法相关挑战、可重复性、所需的专业知识以及每次分析的成本。因此，这篇综述通过提供对可用分析技术的见解，为努力进行临床糖组学分析的研究人员提供了独特的指导。