title: Definitive Screening Design Optimization of Mass Spectrometry Parameters for Sensitive Comparison of Filter and Solid Phase Extraction Purified, INLIGHT Plasma N-Glycans.
abstract: High-throughput, quantitative processing of N-linked glycans would facilitate large-scale studies correlating the glycome with disease and open the field to basic and applied researchers. We sought to meet these goals by coupling filter-aided-N-glycan separation (FANGS) to the individuality normalization when labeling with glycan hydrazide tags (INLIGHT) for analysis of plasma. A quantitative comparison of this method was conducted against solid phase extraction (SPE), a ubiquitous and trusted method for glycan purification. We demonstrate that FANGS-INLIGHT purification was not significantly different from SPE in terms of glycan abundances, variability, functional classes, or molecular weight distributions. Furthermore, to increase the depth of glycome coverage, we executed a definitive screening design of experiments (DOE) to optimize the MS parameters for glycan analyses. We optimized MS parameters across five N-glycan responses using a standard glycan mixture, translated these to plasma and achieved up to a 3-fold increase in ion abundances.
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, United States.
authors.forename: Elizabeth S
authors.initials: ES
authors.lastname: Hecht
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, United States.
authors.forename: James P
authors.initials: JP
authors.lastname: McCord
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, United States.
authors.forename: David C
authors.initials: DC
authors.lastname: Muddiman
mesh_terms: D000818:Animals; D002645:Chickens; D004867:Equipment Design; D013058:Mass Spectrometry / Q000295:instrumentation; D011134:Polysaccharides / Q000097:blood / Q000302:isolation & purification*; D052616:Solid Phase Extraction / Q000295:instrumentation
chemical_list: D011134:Polysaccharides