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亲水作用色谱(HILIC)材料在糖肽的富集与分离中得到越来越广泛的应用.针对目前HILIC材料糖肽选择性不足的缺点,发展新的糖肽富集方法十分必要.本工作发展了一种商品化的弱阳离子交换材料(WCX)在亲水模式下对糖肽的富集方法.首先考察了微固相萃取(SPE)模式下WCX上多肽保留的机理.结果显示,WCX上糖肽的保留同时受乙腈含量和p H的影响.将WCX用于牛胎球蛋白(fetuin)胰蛋白酶酶解液中糖肽的富集,获得了39条糖肽信号.当fetuin与牛血清白蛋白(BSA)酶解液以物质的量比1∶5混合时,富集到26条糖肽信号.富集人免疫球蛋白(Ig G)胰蛋白酶酶解液时获得25条糖肽信号,1∶5时获得23条糖肽信号.实验说明WCX对唾液酸化和非唾液酸化糖肽的富集均有高选择性.该方法拓展了WCX的应用范围,丰富了HILIC材料的种类.
Hydrophilic interaction chromatography (HILIC) materials are more and more widely used in the enrichment and separation of glycopeptides.Aiming at the shortcomings of the lack of glycopeptide selectivity in HILIC, it is necessary to develop new glycopeptide enrichment methods. A method for enriching glycopeptides by a commercially available weak cation exchange material (WCX) was developed.Firstly, the mechanism of peptide retention on WCX in micro-solid-phase extraction (SPE) mode was investigated.The results showed that WCX The retention of glycopeptides was affected by both acetonitrile content and p H.When WCX was used to enrich glycopeptides in fetuin trypsin hydrolyzate, 39 glycopeptide signals were obtained.When fetuin and cow Serum albumin (BSA) hydrolysates were enriched in 26 glycopeptide signals when mixed in a ratio of 1: 5. The enrichment of human immunoglobulin (Ig G) tryptic digests yielded 25 glycopeptides Signal, we obtained 23 glycopeptide signals at 1: 5. The experiment shows that WCX has high selectivity for both sialylated and non-sialylated glycopeptides.This method extends the application range of WCX and enriches the types of HILIC materials.