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URL: https://dx.doi.org/10.17504/protocols.io.ncadase

Authors: Sakari Joenvaara, Mayank Saraswat, Pentti Kuusela, Shruti Saraswat, Rahul Agarwal, Johanna Kaartinen, Asko Jarvinen, Risto Renkonen

Summary: This protocol describes the complete protocol for performing N-Glycoproteomics from plasma samples. Albumin removal is performed prior to protein digestion with trypsin. Tryptic peptide mixture is used for enriching N-glycopeptides by lectin-affinity chromatography using a lectin-mix. Enriched N-glycopeptides are cleaned up by reverse phase chromatogrraphy and analyzed by LC-MS and LC-MS/MS. resulting MS/MS spectra is deconvouted and saved as peak lits file which can be searched with GlycopeptideId software to identify the peptide sequence, glycan composition and proposed glycan struccture. 

Associated Publications: Joenvaara S, Saraswat M, Kuusela P, Saraswat S, Agarwal R, Kaartinen J, Järvinen A, Renkonen R, Quantitative N-glycoproteomics reveals altered glycosylation levels of various plasma proteins in bloodstream infected patients. PLoS ONE 13(3). doi: 10.1371/journal.pone.0195006

Affiliations: University of Helsinki and HUSLAB, University of Helsinki and HUSLAB, Helsinki University Hospital and HUSLAB, University of Helsinki, All India Institute of Medical Sciences, Helsinki University Hospital, Helsinki University Hospital, University of Helsinki and HUSLAB

External URL: https://doi.org/10.1371/journal.pone.0195006

Version: 1

Publication Date: 2018

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