A comparison of N-glycan profiles in human plasma and vitreous fluid
A comparison of N-glycan profiles in human plasma and vitreous fluid
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DOI：
10.1007/s00417-014-2671-x
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发表时间：
2014-06
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通讯作者：
Saori Inafuku;K. Noda;M. Amano;Tetsu Ohashi;Chikako Yoshizawa;W. Saito;Atsuhiro Kanda;S. Nishimura;S. Ishida
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作者：
Saori Inafuku;K. Noda;M. Amano;Tetsu Ohashi;Chikako Yoshizawa;W. Saito;Atsuhiro Kanda;S. Nishimura;S. Ishida
目的探讨非增生性玻璃体视网膜疾病患者血浆和玻璃体标本中n-聚糖的浓度和组成。方法收集11例特发性黄斑孔（MH）患者和9例视网膜前膜（ERM）患者的血浆和玻璃体标本。样品经过酶切预处理，随后从蛋白质中释放的聚糖被捕获在BlotGlyco H珠上。唾液酸被甲基化。处理后的聚糖用氨基功能化肽试剂（aoWR）标记，并从微球中释放，然后用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）进行检测。测定了样品中n -聚糖的浓度和组成。结果玻璃体样品中n -聚糖浓度（132±29 pmol/100 μg蛋白）显著低于血浆样品（714±29 pmol/100 μg蛋白，p< 0.001）。血浆（39.7±1.1%）和玻璃体液（37.2±3.1%）中n -聚糖含量相同，推测其组成为[(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]。相比之下，n -聚糖在玻璃体样品中排名第二（15.6±1.5%），在血浆中排名第七（2.3±0.2%）。结论目前的数据提供了玻璃体液中n -聚糖分布不同于血浆中的有用信息。
PurposeTo investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases.MethodsPlasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). The samples were pretreated for enzymatic cleaving, and subsequently glycans released from proteins were captured on BlotGlyco H beads. Sialic acids were methyl-esterified. Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The concentration and composition of N-glycans in the samples were assessed.ResultsConcentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein,p< 0.001). Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %).ConclusionsThe current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma.