Xin Miaomiao, You Shanshan, Wu Jingyu, Xu Yintai, Li Cheng, Zhu Bojing, Shen Jiechen, Chen Zexuan, Dang Liuyi, Dan Wei, Zhang Xinwen, Sun Shisheng
College of Life Science, Northwest University, Xi'an, Shaanxi Province, 710069, People's Republic of China.
South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, 38925, Vodnany, Czech Republic.
Anal Bioanal Chem. 2022 Dec;414(29-30):8245-8253. doi: 10.1007/s00216-022-04353-4. Epub 2022 Oct 1.
Selecting proper and efficient glycopeptide enrichment approaches are essential for mass spectrometry-based glycoproteomics since glycopeptides are usually with microheterogeneity and low abundance in most biological samples. Herein, we introduced a cotton hydrophilic interaction liquid chromatography (HILIC) approach for large-scale glycopeptide enrichment with 80% acetonitrile/1% trifluoroacetic acid as the optimal sample loading buffer. The comparison of cotton HILIC with Venusil HILIC and mixed anion-exchange (MAX) approaches indicated that cotton HILIC was superior in overall glycopeptide enrichment, whereas Venusil HILIC preferred in complex glycan structures and MAX performed better with high mannose glycans. Exploration of capacity and recovery rate of cotton HILIC illustrated that 5mg cotton packed in a 200μL tip achieved a reasonable glycopeptide enrichment performance (~6% recovery) from ~0.5mg peptides. In conclusion, cotton HILIC can be used as an optional glycopeptide enrichment approach in glycosylation analysis with its specific merit.
由于糖肽在大多数生物样品中通常具有微观异质性且丰度较低，因此选择合适且高效的糖肽富集方法对于基于质谱的糖蛋白质组学至关重要。在此，我们介绍了一种棉花亲水作用液相色谱（HILIC）方法，用于大规模糖肽富集，以80%乙腈/1%三氟乙酸作为最佳上样缓冲液。棉花HILIC与Venusil HILIC和混合阴离子交换（MAX）方法的比较表明，棉花HILIC在整体糖肽富集方面更具优势，而Venusil HILIC在复杂聚糖结构方面表现更佳，MAX在高甘露糖聚糖方面表现更好。对棉花HILIC的容量和回收率的探索表明，填充在200μL尖端的5mg棉花从约0.5mg肽中实现了合理的糖肽富集性能（~6%回收率）。总之，棉花HILIC凭借其独特优点可作为糖基化分析中一种可选的糖肽富集方法。
