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.
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Acknowledgements
We thank B. C. Jansen and K. R. Reiding for in-house developed Python scripts and data analysis support. This work was supported by the European Union (Seventh Framework Programme HighGlycan project, grant number: 278535).
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Holst, S. et al. (2017). High-Throughput and High-Sensitivity Mass Spectrometry-Based N-Glycomics of Mammalian Cells. In: Lauc, G., Wuhrer, M. (eds) High-Throughput Glycomics and Glycoproteomics. Methods in Molecular Biology, vol 1503. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6493-2_14
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DOI: https://doi.org/10.1007/978-1-4939-6493-2_14
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