整体柱用于糖蛋白质分离富集研究新进展

Recent Advances in Monolithic Columns and Their Application in Separation and Enrichment of Glycoproteins

蛋白质糖基化是最常见、最重要的蛋白质翻译后修饰之一,由糖链和多肽链以多种形式共价修饰而成。糖蛋白在生物体内种类繁多,分布广泛,具有重要的生理功能。复杂生物体系中糖蛋白的绝对丰度低,高丰度非糖蛋白质的存在对低丰度糖蛋白质产生夹带、包覆以至掩盖的作用,以现有的技术难以发现并分离鉴定大多数低丰度糖蛋白,已成为当前糖蛋白质组学研究面临的瓶颈之一。去除高丰度非糖蛋白质或富集低丰度的糖蛋白是解决这一问题的有效途径。整体柱是一种通过有机或无机聚合方法在柱内进行原位聚合的连续床固定相,具有制备简单,多孔性、柱容量高,能实现快速、高效分离等特点。整体柱制备技术的发展为糖蛋白质的分离富集研究提供了一种新途径。该文简要介绍了整体柱的制备方法和分类,综述了包括凝集素亲和整体柱、苯硼酸型整体柱和固定金属离子化在内的整体柱在糖蛋白分离富集方面的最新研究进展,并对其应用前景进行了展望。引用文献65篇。

With the accomplishment of the gene sequencing of Human Genomic Project（HGP）,increasing attention is focusing on proteomic research.Glycosylation of proteins is the most common and important post-translational modification,which is composed of many covalent forms of polypeptide and oligosaccharide.The structure of sugar chain plays a crucial role in various biological processes.However,due to the existence of high abundant non-glycoproteins,the low abundant glycoproteins were prone to be embedded and covered by nonglycoproteins and thus causing serious interference in further identification.The existing technologies are difficult to isolate and detect low abundant glycoproteins from complex matrices,which has become the one of the major bottlenecks currently encountered in glycoproteomics.Monolithic column,as an integrated continuous porous stationary phase,was prepared by in-situ polymerization or sol-gel process inside the capillary,which can be taken as an alternative approach for isolation and enrichment of glycoproteins due to the merits of simple preparation procedure,high porosity,high capacity and high efficient separation.In this paper,the principle,preparation and classification of monolithic columns were briefly introduced.The recent advances of monolithic columns in selective separation and enrichment of glycoproteins were summarized,which included lectin-based affinity monoliths,boronate affinity monoliths and immobilized metal affinity monoliths.In addition,the prospect of monolithic column for enrichment of glycoproteins was also presented.Totally,65 references were reviewed.