High-Throughput and High-Sensitivity Mass Spectrometry-Based N-Glycomics of Mammalian Cells

Stephanie Holst, Gabi W van Pelt, Wilma E Mesker, Rob A Tollenaar, Ana I Belo, Irma van Die, Yoann Rombouts, Manfred Wuhrer

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

The current protocols for glycomic analysis of cells often require a large quantity of material (5-20 million cells). In order to analyze the N-glycosylation from small amounts of cells (≤1 million) as obtained from, for example, primary cell lines or cell sorting, and in a higher throughput approach, we set up a robust 96-well format PVDF-membrane based N-glycan release protocol followed by linkage-specific sialic acid stabilization, cleanup, and MALDI-TOF-MS analysis. We further evaluated the influence of PNGase F incubation time on the N-glycan profile.

Original languageEnglish
Pages (from-to)185-196
Number of pages12
JournalMethods in Molecular Biology
Volume1503
DOIs
Publication statusPublished - 2017

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