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Glycan‐Selective MALDI In‐Source Decay Analysis of Intact Glycoproteins

Glycan‐Selective MALDI In‐Source Decay Analysis of Intact Glycoproteins A simple matrix‐assisted laser desorption/ionization‐time‐of‐flight (MALDI‐TOF) mass spectrometry (MS) approach has been developed to analyze the structural pattern of glycan on glycoproteins without any digestive or modification pretreatment. For a 7 pmol sample of intact glycoprotein, RNase B, a solid ionic matrix DHB‐aniline‐sodium enabled a regioselective in‐source decay (ISD) and glycan‐selective ionization as a sodium adduct ion, [M+Na]+. This matrix significantly reduced the formation of peptide‐fragment‐derived ions, and a high‐mannose‐type glycan pattern was observed. This ISD product ion was formed by selective cleavage of the reducing end of the glycan by 0,2A or 2,4A type cross‐ring cleavage. Pseudo‐MS3‐type post‐source decay (PSD) analysis of an A type ion indicated the regioselectivity of the ISD fragmentation and the glycan sequence. Combining ISD glycomics with conventional ISD proteomics will accelerate quality control for glycoprotein drugs and biomarker discovery. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Analysis & Sensing Wiley

Glycan‐Selective MALDI In‐Source Decay Analysis of Intact Glycoproteins

Analysis & Sensing , Volume 2 (2) – Mar 1, 2022

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Publisher
Wiley
Copyright
© 2022 Wiley‐VCH GmbH
eISSN
2629-2742
DOI
10.1002/anse.202100040
Publisher site
See Article on Publisher Site

Abstract

A simple matrix‐assisted laser desorption/ionization‐time‐of‐flight (MALDI‐TOF) mass spectrometry (MS) approach has been developed to analyze the structural pattern of glycan on glycoproteins without any digestive or modification pretreatment. For a 7 pmol sample of intact glycoprotein, RNase B, a solid ionic matrix DHB‐aniline‐sodium enabled a regioselective in‐source decay (ISD) and glycan‐selective ionization as a sodium adduct ion, [M+Na]+. This matrix significantly reduced the formation of peptide‐fragment‐derived ions, and a high‐mannose‐type glycan pattern was observed. This ISD product ion was formed by selective cleavage of the reducing end of the glycan by 0,2A or 2,4A type cross‐ring cleavage. Pseudo‐MS3‐type post‐source decay (PSD) analysis of an A type ion indicated the regioselectivity of the ISD fragmentation and the glycan sequence. Combining ISD glycomics with conventional ISD proteomics will accelerate quality control for glycoprotein drugs and biomarker discovery.

Journal

Analysis & SensingWiley

Published: Mar 1, 2022

Keywords: glycomics; glycoproteins; in-source decay; MALDI; solid ionic matrices

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