Franz Andreas H, Ta Nathan, Nguyen Ella, Bromley Kendall, Rosenblatt Hana, Fabro Aprilrose, Lin-Cereghino Geoff
Department of Chemistry, University of the Pacific, 3601 Pacific Avenue, Stockton, CA, 95211, USA.
Department of Biology, University of the Pacific, 3601 Pacific Avenue, Stockton, CA, 95211, USA.
Anal Sci. 2025 Mar;41(3):297-310. doi: 10.1007/s44211-024-00700-w. Epub 2024 Dec 10.
Oligosaccharides were covalently labeled with 1-pyrenemethylamine (1-PMA) in 15 min at 75 °C with high yield, separated by quantitative HPLC in less than 20 min, and characterized by off-line Matrix-Assisted Laser Desorption/Ionization (MALDI) Time-of-Flight (TOF) mass spectrometry (MS). A new MALDI mass spectral precursor ion for fragmentation studies was observed from 2,5-dihydroxybenzoic acid matrix (DHB, hydroquinone/benzoquinone redox system) through photo-induced reductive elimination. The resulting high-energy protonated glycosylamine provided excellent fragmentation efficiency (Y-, B-ions, and combination losses), with ions covering almost the entire structure of several oligosaccharides at the low picomol level. With the new method, glycans from commercially available standard glycoproteins and glycans from bioengineered β-lactoglobulin expressed in the bgs13 mutant of Pichia pastoris were analyzed.
寡糖在75℃下与1-芘甲胺（1-PMA）共价标记15分钟，产率高，在不到20分钟内通过定量高效液相色谱分离，并用离线基质辅助激光解吸/电离（MALDI）飞行时间（TOF）质谱（MS）进行表征。通过光诱导还原消除，从2,5-二羟基苯甲酸基质（DHB，对苯二酚/苯醌氧化还原体系）中观察到一种用于碎片研究的新的MALDI质谱前体离子。所得的高能质子化糖基胺具有出色的碎片效率（Y-离子、B-离子和组合损失），在低皮摩尔水平下，离子几乎覆盖了几种寡糖的整个结构。采用这种新方法，对市售标准糖蛋白中的聚糖以及在毕赤酵母bgs13突变体中表达的生物工程β-乳球蛋白中的聚糖进行了分析。
