Post-Column Make-Up Flow (PCMF) Enhances the Performance of Capillary-Flow PGC-LC-MS/MS-Based Glycomics

Analytical Chemistry
Hannes HinneburgMorten Thaysen-Andersen

Abstract

Deep characterization of biologically relevant glycans remains challenging. Porous graphitized carbon-liquid chromatography tandem mass spectrometry (PGC-LC-MS/MS) enables the quantitative elucidation of glycan fine structures. However, the early PGC-LC elution of smaller glycans (tri-, tetra-, and pentasaccharides) at low organic solvent content hampers their detection. In efforts to improve the glycan profiling sensitivity and accuracy, we present a new capillary-flow PGC-LC-MS/MS-based configuration comprising a post-column make-up flow (PCMF) that supplies an ion-promoting organic solvent to separated glycans prior to their detection by MS. The analytical performance of this setup was systematically evaluated against our existing capillary-flow PGC-LC-MS/MS platform (Jensen et al., Nat. Protoc. 2012, 7, 1299). Specifically, the ion intensities and signal-to-noise ratios of various classes of nonderivatized glycans from N- and O-glycoproteins and fructooligosaccharide mixtures were compared using methanol (MeOH)-, isopropanol (IPA)-, and acetonitrile (ACN)-based PCMF at various concentrations. In particular, ACN- and IPA-based PCMF dramatically increased the signal response across all glycan types (30- to 100-fold), improved...Continue Reading

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Citations

Aug 15, 2019·Biological Reviews of the Cambridge Philosophical Society·Harry C TjondroMorten Thaysen-Andersen
Jul 16, 2020·The Journal of Biological Chemistry·Vignesh VenkatakrishnanAnna Karlsson-Bengtsson
Sep 10, 2020·Analytical Methods : Advancing Methods and Applications·Katherine L Sanders, James L Edwards
Aug 17, 2019·Proteomics·Sayantani ChatterjeeMorten Thaysen-Andersen
Mar 10, 2020·Glycobiology·Hannes HinneburgMorten Thaysen-Andersen
Jan 14, 2021·Biochemical Society Transactions·Anastasia ChernykhMorten Thaysen-Andersen
Dec 5, 2020·The Journal of Biological Chemistry·Harry C TjondroMorten Thaysen-Andersen
Feb 5, 2021·Journal of Clinical Medicine·Sayantani ChatterjeeMorten Thaysen-Andersen
Aug 28, 2020·Molecular Aspects of Medicine·Julian UgonottiMorten Thaysen-Andersen
Sep 3, 2020·Journal of Proteome Research·Rongxia LanShisheng Sun
Apr 30, 2021·Analytical Chemistry·Katarina MadunićGuinevere S M Lageveen-Kammeijer
May 23, 2020·Analytical Chemistry·Miloslav SandaRadoslav Goldman
Sep 9, 2021·Mass Spectrometry Reviews·Guinevere S M Lageveen-KammeijerManfred Wuhrer

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