Integrated Mass Spectrometry-based Analysis of Plasma Glycoproteins and Their Glycan Modifications

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2011 Mar 5

PMID 21372808

Citations 11

Authors

Hong Wang

Chee-Hong Wong

Alice Chin

Ayumu Taguchi

Allen Taylor

Samir Hanash

Sadanori Sekiya

Hidenori Takahashi

Masaki Murase

Shigeki Kajihara

Shinichi Iwamoto

Koichi Tanaka

Affiliations

Soon will be listed here.

Abstract

We present a protocol for the identification of glycosylated proteins in plasma followed by elucidation of their individual glycan compositions. The study of glycoproteins by mass spectrometry is usually based on cleavage of glycans followed by separate analysis of glycans and deglycosylated proteins, which limits the ability to derive glycan compositions for individual glycoproteins. The methodology described here consists of 2D HPLC fractionation of intact proteins and liquid chromatography-multistage tandem mass spectrometry (LC-MS/MS(n)) analysis of digested protein fractions. Protein samples are separated by 1D anion-exchange chromatography (AEX) with an eight-step salt elution. Protein fractions from each of the eight AEX elution steps are transferred onto the 2D reversed-phase column to further separate proteins. A digital ion trap mass spectrometer with a wide mass range is then used for LC-MS/MS(n) analysis of intact glycopeptides from the 2D HPLC fractions. Both peptide and oligosaccharide compositions are revealed by analysis of the ion fragmentation patterns of glycopeptides with an intact glycopeptide analysis pipeline.

Citing Articles

Plasma/Serum Proteomics based on Mass Spectrometry.

Zhu Y Protein Pept Lett. 2024; 31(3):192-208.

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Rational Development of Hypervalent Glycan Shield-Binding Nanoparticles with Broad-Spectrum Inhibition against Fatal Viruses Including SARS-CoV-2 Variants.

Li Y, Xu S, Ye Q, Chi H, Guo Z, Chen J Adv Sci (Weinh). 2022; 10(2):e2202689.

PMID: 36377484 PMC: 9839850. DOI: 10.1002/advs.202202689.

Microbowls with Controlled Concavity for Accurate Microscale Mass Spectrometry.

Xu L, Li X, Li W, Chang K, Yang H, Tao N Adv Mater. 2022; 34(12):e2108194.

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N-Linked Glycopeptide Identification Based on Open Mass Spectral Library Search.

An Z, Shu Q, Lv H, Shu L, Wang J, Yang F Biomed Res Int. 2018; 2018:1564136.

PMID: 30186849 PMC: 6112209. DOI: 10.1155/2018/1564136.

Data-independent oxonium ion profiling of multi-glycosylated biotherapeutics.

Madsen J, Farutin V, Lin Y, Smith S, Capila I MAbs. 2018; 10(7):968-978.

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