糖组学,在基因组学和蛋白质组学之后发展起来的一种新兴的“组学”技术,是一门研究构图的学科,结构,和细胞中的糖的功能,组织,和有机体。糖组学在理解主要生命活动的规律中起着关键作用,疾病的预防和治疗,以及药品质量控制和开发。目前,聚糖的结构分析主要依赖于质谱。然而,聚糖在生物样品中具有低丰度。此外,因素,如可变的单糖组成,糖苷键位置和模式的差异,不同的分支结构,导致聚糖的组成和结构的复杂性,给糖组学研究带来巨大挑战。液相色谱可以有效去除基质干扰并增强聚糖分离以改善聚糖的质谱响应。因此,液相色谱和液相色谱-质谱联用技术是解决这些问题的重要技术手段,这些技术在糖组学研究中发挥着不可或缺的作用。不同的研究突出了各种类型液相色谱的应用的异同,这也反映了这项技术的多功能性和灵活性。在这次审查中,我们首先从色谱分离机理的角度讨论了聚糖的富集方法及其在糖组学研究中的应用。然后我们比较了这些方法的优缺点。一些聚糖富集模式包括亲和力,亲水相互作用,尺寸排除,和多孔石墨化碳吸附。许多新开发的材料表现出优异的聚糖富集能力。我们列举了反相高效液相色谱(RP-HPLC)的分离机理,高效阴离子交换色谱(HPAEC),亲水相互作用色谱(HILIC),和多孔石墨碳(PGC)色谱在聚糖的分离和分析中,并描述了这些方法在分离聚糖中的应用,糖缀合物,和糖衍生物。在这些方法中,HILIC和PGC色谱法是应用最广泛的,而HPAEC和RP-HPLC较不常用。HILIC和RP-HPLC模式通常用于分离衍生的聚糖。衍生化后聚糖的电离效率和可检测性显著提高。然而,衍生化过程相对繁琐,和副产物不可避免地影响检测结果的准确性和完整性。HPAEC和PGC色谱对非衍生聚糖具有良好的分离效果,但与低丰度聚糖检测完整性相关的问题仍然存在,因为它们的检测效果不佳.因此,必须选择特定样品或目标分析物的适当分析方法或相互验证。最后,我们重点介绍了各种色谱-质谱联用技术在糖组学分析中的研究进展。近年来,由于色谱分离技术的发展,糖组学研究取得了重大进展。然而,仍然存在一些重大挑战。随着新型分离材料和方法的不断发展,色谱技术有望在未来的糖组学研究中发挥关键作用。
Glycomics, an emerging \"omics\" technology that was developed after genomics and proteomics, is a discipline that studies the composition, structure, and functions of glycomes in cells, tissues, and organisms.
Glycomics plays key roles in understanding the laws of major life activities, disease prevention and treatment, and drug quality control and development. At present, the structural analysis of glycans relies mainly on mass spectrometry. However, glycans have low abundance in biological samples. In addition, factors such as variable monosaccharide compositions, differences in glycosidic bond positions and modes, diverse branching structures, contribute to the complexity of the compositions and structures of glycans, posing great challenges to glycomics research. Liquid chromatography can effectively remove matrix interferences and enhance glycan separation to improve the mass spectrometric response of glycans. Thus, liquid chromatography and liquid chromatography coupled with mass spectrometry are important technical tools that have been actively applied to solve these problems; these technologies play indispensable roles in
glycomics research. Different studies have highlighted similarities and differences in the applications of various types of liquid chromatography, which also reflects the versatility and flexibility of this technology. In this review, we first discuss the enrichment methods for glycans and their applications in
glycomics research from the perspective of chromatographic separation mechanisms. We then compare the advantages and disadvantages of these methods. Some glycan-enrichment modes include affinity, hydrophilic interactions, size exclusion, and porous graphitized carbon adsorption. A number of newly developed materials exhibit excellent glycan-enrichment ability. We enumerate the separation mechanisms of reversed-phase high performance liquid chromatography (RP-HPLC), high performance anion-exchange chromatography (HPAEC), hydrophilic interaction chromatography (HILIC), and porous graphitic carbon (PGC) chromatography in the separation and analysis of glycans, and describe the applications of these methods in the separation of glycans, glycoconjugates, and glyco-derivatives. Among these methods, HILIC and PGC chromatography are the most widely used, whereas HPAEC and RP-HPLC are less commonly used. The HILIC and RP-HPLC modes are often used for the separation of derived glycans. The ionization efficiency and detectability of glycans are significantly improved after derivatization. However, the derivatization process is relatively cumbersome, and byproducts inevitably affect the accuracy and completeness of the detection results. HPAEC and PGC chromatography exhibit good separation effects on nonderivative glycans, but issues related to the detection integrity of low-abundance glycans owing to their poor detection effect continue to persist. Therefore, the appropriate analytical method for a specific sample or target analyte or mutual verification must be selected. Finally, we highlight the research progress in various chromatographic methods coupled with mass spectrometry for glycomics analysis. Significant progress has been made in
glycomics research in recent years owing to advancements in the development of chromatographic separation techniques. However, several significant challenges remain. As the development of novel separation materials and methods continues, chromatographic techniques may be expected to play a critical role in future glycomics research.