Profile of native N-linked glycan structures from human serum using high performance liquid chromatography on a microfluidic chip and time-of-flight mass spectrometry.
Profile of native N-linked glycan structures from human serum using high performance liquid chromatography on a microfluidic chip and time-of-flight mass spectrometry.
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在微流体芯片和飞行时间质谱上使用高性能液相色谱法的天然N连接聚糖结构的剖面。
DOI：
10.1002/pmic.200800249
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发表时间：
2009-04
期刊：
影响因子：
3.4
通讯作者：
Lebrilla, Carlito B.
中科院分区：
文献类型：
--
作者：
Chu, Caroline S.;Ninonuevo, Milady R.;Clowers, Brian H.;Perkins, Patrick D.;An, Hyun Joo;Yin, Hongfeng;Killeen, Kevin;Miyamoto, Suzanne;Grimm, Rudolf;Lebrilla, Carlito B.
蛋白质糖基化包括在不需要糖链模板的分步过程中加入单糖。因此，鉴定包括异构体在内的众多糖形式，有助于阐明特定多糖的生物学功能(S)。建立了一种利用在线纳米液色谱(NanoLC)和高分辨飞行时间质谱仪(TMS)对人血清中N-连接低聚糖进行非衍生化释放的分析方法。用基质辅助激光解吸电离傅立叶变换离子回旋共振质谱仪(MALDI FT-ICR MS)和微芯片液相色谱质谱(HPLC-Chip/TOF MS)对N-连接的低聚糖进行了分析。采用两种微流控芯片，糖链芯片(40nL富集柱，43×0.075 mm ID分析柱)和高容量芯片(160nL富集柱，140×0.075 mm ID分析柱)，均以石墨化碳为固定相。这两种芯片在分离注射之间的中性和阴离子低聚糖的异相混合物方面都提供了良好的灵敏度和重现性。增加富集柱和分析柱的长度和体积可以提高峰的分辨率。人血清中含量最丰富的低聚糖为复合型N-连接低聚糖，约占总糖链的96%，杂交型和高甘露糖型低聚糖占其余的4%。
Protein glycosylation involves the addition of monosaccharides in a stepwise process requiring no glycan template. Therefore, identifying the numerous glycoforms, including isomers, can help elucidate the biological function(s) of particular glycans. A method to assess the diversity of the N-linked oligosaccharides released from human serum without derivatization has been developed using on-line nano-liquid chromatography (nanoLC) and high resolution time-of-flight mass spectrometry. The N-linked oligosaccharides were analyzed with matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI FT-ICR MS) and microchip liquid chromatography mass spectrometry (HPLC-Chip/TOF MS). Two microfluidic chips were employed, the glycan chip (40 nL enrichment column, 43×0.075 mm ID analytical column) and the high capacity chip (160 nL enrichment column, 140×0.075 mm ID analytical column), both with graphitized carbon as the stationary phase. Both chips offered good sensitivity and reproducibility in separating a heterogeneous mixture of neutral and anionic oligosaccharides between injections. Increasing the length and volume of the enrichment and the analytical columns improved resolution of the peaks. Complex type N-linked oligosaccharides were the most abundant oligosaccharides in human serum accounting for ~96% of the total glycans identified, while hybrid and high mannose type oligosaccharides comprise the remaining ~4%.
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影响因子：
4.3
作者：
Butler, M;Quelhas, D;Rudd, PM
通讯作者：
Rudd, PM
影响因子：
7
作者：
Kirmiz, Crystal;Li, Bensheng;Miyamoto, Suzanne
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Miyamoto, Suzanne
影响因子：
4.4
作者：
Kyselova, Zuzana;Mechref, Yehia;Novotny, Milos V.
通讯作者：
Novotny, Milos V.
影响因子：
6.1
作者：
Ninonuevo, Milady R.;Park, Youmie;Lebrilla, Carlito B.
通讯作者：
Lebrilla, Carlito B.
影响因子：
4.1
作者：
Itoh, S;Kawasaki, N;Hayakawa, T
通讯作者：
Hayakawa, T