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Desalting paper spay mass spectrometry (DPS-MS) for rapid detection of glycans and glycoconjugates
International Journal of Mass Spectrometry ( IF 1.8 ) Pub Date : 2021-08-28 , DOI: 10.1016/j.ijms.2021.116688
Kai-Yuan Chiu 1 , Qi Wang 1 , Harsha P Gunawardena 2 , Michael Held 3, 4 , Ahmed Faik 4, 5 , Hao Chen 1
Affiliation  

The detection of glycans and glycoconjugates has gained increasing attention in biological fields. Traditional mass spectrometry (MS)-based methods for glycoconjugate analysis are challenged with poor sensitivity when dealing with complex biological samples. We developed a desalting paper spray mass spectrometry (DPS-MS) strategy to overcome the issue of signal suppression of carbohydrates in salted buffers. Glycans and glycoconjugates (i.e., glycopeptides, nucleotide sugars, etc.) in non-volatile buffer (e.g., Tris buffer) can be loaded on the paper substrate from which buffers can be removed by washing with ACN/H2O (90/10 v/v) solvent. Glycans or glycoconjugates can then be eluted and spray ionized by adding ACN/H2O/formic acid (FA) (10/90/1 v/v/v) solvent and applying a high voltage (HV) to the paper substrate. Our results showed that DPS-MS is applicable for direct detection of intact glycopeptides and nucleotide sugars as well as determination of glycosylation profiling of antibody, such as NIST monoclonal antibody IgG (NISTmAb). NISTmAb was deglycosylated with PNGase F to release N-linked oligosaccharides. Twenty-six N-linked oligosaccharides were detected by DPS-MS within a 5-min timeframe without the need for enrichment or derivatization. This work demonstrates that DPS-MS allows fast and sensitive detection of glycans/oligosaccharides and glycosylated species in complex matrices and has great potential in bioanalysis.



中文翻译:

脱盐纸喷雾质谱 (DPS-MS) 用于快速检测聚糖和糖缀合物

聚糖和糖缀合物的检测在生物学领域越来越受到关注。在处理复杂的生物样品时,传统的基于质谱 (MS) 的糖缀合物分析方法面临着灵敏度低的挑战。我们开发了一种脱盐纸喷雾质谱 (DPS-MS) 策略来克服盐缓冲液中碳水化合物的信号抑制问题。非挥发性缓冲液(例如,Tris 缓冲液)中的聚糖和糖缀合物(即,糖肽、核苷酸糖)可以加载到纸基材上,可以通过用 ACN/H 2 O (90/10 ) 洗涤从纸基材上去除缓冲液v/v) 溶剂。然后可以通过添加 ACN/H 2洗脱和喷雾离子化聚糖或糖缀合物O/甲酸 (FA) (10/90/1 v/v/v) 溶剂并在纸基材上施加高压 (HV)。我们的结果表明,DPS-MS 适用于直接检测完整的糖肽和核苷酸糖以及测定抗体的糖基化谱,例如 NIST 单克隆抗体 IgG (NISTmAb)。NISTmAb 用 PNGase F 去糖基化以释放 N-连接寡糖。DPS-MS 在 5 分钟内检测到 26 种 N 连接寡糖,无需富集或衍生化。这项工作表明,DPS-MS 可以快速、灵敏地检测复杂基质中的聚糖/寡糖和糖基化物质,在生物分析中具有巨大潜力。

更新日期:2021-09-07
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