Integration of click maltose hydrophilic interaction chromatography, SCX preolumn and PNGase F immobilized enzymatic reactor for N-linked glycosylation sites profiling

	Integration of click maltose hydrophilic interaction chromatography, SCX preolumn and PNGase F immobilized enzymatic reactor for N-linked glycosylation sites profiling
	Qu YY(曲焱焱) ; Yuan HM(袁辉明) ; Zhang LH(张丽华) ; Liang Z(梁振) ; Zhang YK(张玉奎)
	2011
会议名称	26th international symposium on microscale bioseparations
会议日期	2011-5-1
会议地点	圣地亚哥
页码	75-0
通讯作者	张丽华
中文摘要	n-glycosylation is one of the most common and complex post-translational modifications of proteins, and plays an important role in many cellular interactions and pathogenesis of diseases. global mapping of n-linked glycosylation sites is a prerequisite for fully understanding the biological functions of n-linked glycoproteins. therefore, the development of a fast and specific strategy to achieve high-throughput and high-sensitivity glycoproteome analysis becomes imperative. the traditional n-linked glycosylation sites profiling is usually off-line performed, with drawbacks such as long analysis time, resultant sample loss and manual manipulation. in our recent work, an integrated sample pretreatment system, composed of a click maltose hydrophilic interaction chromatography (hilic) column, a strong cation exchange (scx) precolumn and a pngase f immobilized enzymatic reactor (imer), was established for the simultaneous glycopeptide enrichment, sample buffer exchange, and on-line deglycosylation. compared with the conventional off-line method, the analysis time, including glycopeptide enrichment and deglycosylation was shortened to 1 h, and the detection limit as low as 5 fmol was achieved. moreover, such a system was successfully applied for analyzing the digest of soluble fraction extracted from rat brain. a total of 120 unique glycoprotein groups and 196 n-linked glycosylation sites were identified, with the injected digests amount as 6 µg. all these results demonstrated that the integrated system is of great promise to achieve fast and high sensitive n-linked glycosylation sites profiling, which could be further online coupled with nano hplc-esi/ms/ms to achieve high-throughput glycoproteome analysis.
合作状况	墙报
会议主办者	casss
会议录	26th international symposium on microscale bioseparations
会议录出版者	待补充
会议录出版地	待补充
学科主题	分析化学
语种	英语
内容类型	会议论文
源URL	[http://159.226.238.44/handle/321008/116025]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
推荐引用方式 GB/T 7714	Qu YY,Yuan HM,Zhang LH,et al. Integration of click maltose hydrophilic interaction chromatography, SCX preolumn and PNGase F immobilized enzymatic reactor for N-linked glycosylation sites profiling[C]. 见:26th international symposium on microscale bioseparations. 圣地亚哥. 2011-5-1.