Ion Mobility Mass Spectrometry for Ion Recovery and Clean-Up of MS and MS/MS Spectra Obtained from Low Abundance Viral Samples
Ion Mobility Mass Spectrometry for Ion Recovery and Clean-Up of MS and MS/MS Spectra Obtained from Low Abundance Viral Samples
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DOI：
10.1007/s13361-015-1163-5
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发表时间：
2015-10-01
影响因子：
3.2
通讯作者：
Scrivens, Jim H.
中科院分区：
文献类型：
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作者：
Harvey, David J.;Crispin, Max;Scrivens, Jim H.
从少量物质（例如来自病毒的糖蛋白）释放的N-聚糖的复杂混合物的许多样品对于质谱分析存在问题，因为存在难以通过常规方法去除而不涉及样品损失的污染物质。本研究描述了使用离子迁移率从此类样品中提取聚糖谱，并在目标m/z值从目标化合物中捕获其他离子时获得干净的CID谱。N-聚糖从SDS-PAGE凝胶内酶促释放，从人免疫缺陷病毒的代表性重组糖蛋白gp 120释放，并通过负离子模式下的直接输注电喷雾，然后用沃茨Synapt G2质谱仪（沃茨MS-Technologies，Manchester，UK）进行离子迁移率检查。在原始电喷雾光谱由于样品浓度低或存在污染而仅显示少数聚糖离子的情况下，从样品中获得了单电荷、双电荷和三电荷N-聚糖的清洁图谱。当聚糖的分子离子显示与来自具有相似m/z值的碎片或多电荷离子的离子重合时，离子迁移率也使得能够从聚糖获得未污染的CID光谱。这种技术被证明是非常宝贵的，从许多CID光谱去除外来离子。这种离子的存在通常会产生难以解释的光谱。大多数CID光谱，即使是那些丰富的聚糖成分，受益于这样的清理，表明离子迁移率提供的额外维度是非常宝贵的这种类型的研究。
Many samples of complex mixtures of N-glycans released from small amounts of material, such as glycoproteins from viruses, present problems for mass spectrometric analysis because of the presence of contaminating material that is difficult to remove by conventional methods without involving sample loss. This study describes the use of ion mobility for extraction of glycan profiles from such samples and for obtaining clean CID spectra when targeted m/z values capture additional ions from those of the target compound. N-glycans were released enzymatically from within SDS-PAGE gels, from the representative recombinant glycoprotein, gp120 of the human immunodeficiency virus, and examined by direct infusion electrospray in negative mode followed by ion mobility with a Waters Synapt G2 mass spectrometer (Waters MS-Technologies, Manchester, UK). Clean profiles of singly, doubly, and triply charged N-glycans were obtained from samples in cases where the raw electrospray spectra displayed only a few glycan ions as the result of low sample concentration or the presence of contamination. Ion mobility also enabled uncontaminated CID spectra to be obtained from glycans when their molecular ions displayed coincidence with ions from fragments or multiply charged ions with similar m/z values. This technique proved to be invaluable for removing extraneous ions from many CID spectra. The presence of such ions often produces spectra that are difficult to interpret. Most CID spectra, even those from abundant glycan constituents, benefited from such clean-up, showing that the extra dimension provided by ion mobility was invaluable for studies of this type.