Book Chapter10.1007/978-1-59745-022-5_4
Solid-phase permethylation for glycomic analysis.
78
TL;DR: This chapter discusses in detail a miniaturized version of the widely used permethylation technique which permits quantitative derivatization of oligosaccharides derived from minute quantities of glycoprotein through packing of sodium hydroxide powder or beads in a microcolumn format.
read more
Abstract: This chapter discusses in detail a miniaturized version of the widely used permethylation technique which permits quantitative derivatization of oligosaccharides derived from minute quantities of glycoprotein. The approach involves packing of sodium hydroxide powder or beads in a microcolumn format, including spin columns, fused silica capillaries (500 microm i.d.) and plastic tubes (1 mm i.d.). The derivatization proceeds effectively in less than a minute time scale and it is applicable to glycans derived from femtomole quantities of glycoproteins. Prior to mass spectrometry (MS), methyl iodide is added to analytes suspended in dimethyl sulfoxide solution containing traces of water. The reaction mixture is then immediately infused through the microreactor. The packed sodium hydroxide powder or beads inside the microcolumns minimize oxidative degradation and peeling reactions which are otherwise commonly associated with the conventional permethylation technique. In addition, this solid-phase permethylation approach eliminates the need for excessive sample clean-up. As demonstrated below, picomole amounts of various types of glycans derived from model glycoproteins as well as real samples, including linear and branched, sialylated and neutral glycans were shown to become rapidly and efficiently permethylated through this approach.
read more
Chat with Paper
AI Agents for this Paper
Find similar papers on Google Scholar, PubMed and Arxiv
Write a critical review of this paper
Analyze citations of this paper to find unaddressed research gaps
Citations
Effective use of mass spectrometry for glycan and glycopeptide structural analysis.
Nancy Leymarie,Joseph Zaia +1 more
TL;DR: Mass spectrometry methods are used for mapping glycoprotein glycosylation and detailed glycan structural determination and enables precise characterization of recombinant glycoproteins in the pharmaceutical industry and academic biomedicine.
196
Matrix assisted laser desorption ionization imaging mass spectrometry workflow for spatial profiling analysis of N-linked glycan expression in tissues.
Thomas W. Powers,E. Ellen Jones,Lucy Betesh,Patrick R. Romano,Peng Gao,John A. Copland,Anand Mehta,Richard R. Drake +7 more
TL;DR: The ability to differentially profile N-glycans and correlate their molecular expression to histopathological changes can offer new approaches to identifying novel disease related targets for biomarker and therapeutic applications.
168
Characterization of isomeric glycan structures by LC-MS/MS.
TL;DR: Recent advances in porous graphitized carbon, reverse phase, ion exchange, and hydrophilic interaction LC utilized in conjunction with MS, for the characterization of protein glycosylation are described with an emphasis on methods capable of resolving isomeric glycan structures.
147
Identifying cancer biomarkers by mass spectrometry-based glycomics
TL;DR: This review focuses on describing, discussing, and evaluating the different Glycomic methods employed to characterize and quantify glycomic changes associated with cancers of different organs, including breast, colon, esophagus, liver, ovarian, pancreas, and prostate.
Direct comparison of derivatization strategies for LC-MS/MS analysis of N-glycans
TL;DR: Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans.
References
A simple and rapid method for the permethylation of carbohydrates
Ionel Ciucanu,Francisc Kerek +1 more
TL;DR: In this paper, a new method for the permethylation of sugars involving methyl iodide, a solid base (NaOH, KOH, or tert-BuOH/NaOH), and methyl sulphoxide was suggested.
3.5K
Solid-phase permethylation of glycans for mass spectrometric analysis
TL;DR: A miniaturized approach was developed for quantitative permethylation of oligosaccharides, which involves packing of sodium hydroxide powder in microspin columns or fused-silica capillaries, permitting effective derivatization in less than a minute at microscale.
Elimination of oxidative degradation during the per-O-methylation of carbohydrates.
TL;DR: Evidence is presented that this process occurs only under anhydrous conditions when there is a long reaction time between the carbohydrate dissolved in dimethyl sulfoxide and methyl iodide, followed by reaction with the base.
257
Microscale nonreductive release of O-linked glycans for subsequent analysis through MALDI mass spectrometry and capillary electrophoresis.
TL;DR: A new beta-elimination-based procedure has been devised for a microscale release of O-linked oligosaccharides from glycoproteins that renders the reducing end intact and permits us to investigate structurally O- linked oligosACcharides at very low levels.
224
Structural Characterization of Carbohydrate Sequence, Linkage, and Branching in a Quadrupole Ion Trap Mass Spectrometer: Neutral Oligosaccharides and N-Linked Glycans
TL;DR: Several oligosaccharide and N-linked glycan samples have been utilized to evaluate structural detail obtained with an ion trap mass spectrometer (ITMS), and this technology appears to provide a significant step toward the goal of characterizing all aspects of carbohydrate structure using a single instrument.
166