title: Ion Mobility Mass Spectrometry for Ion Recovery and Clean-Up of MS and MS/MS Spectra Obtained from Low Abundance Viral Samples.
abstract: Many samples of complex mixtures of N-glycans released from small amounts of material, such as glycoproteins from viruses, present problems for mass spectrometric analysis because of the presence of contaminating material that is difficult to remove by conventional methods without involving sample loss. This study describes the use of ion mobility for extraction of glycan profiles from such samples and for obtaining clean CID spectra when targeted m/z values capture additional ions from those of the target compound. N-glycans were released enzymatically from within SDS-PAGE gels, from the representative recombinant glycoprotein, gp120 of the human immunodeficiency virus, and examined by direct infusion electrospray in negative mode followed by ion mobility with a Waters Synapt G2 mass spectrometer (Waters MS-Technologies, Manchester, UK). Clean profiles of singly, doubly, and triply charged N-glycans were obtained from samples in cases where the raw electrospray spectra displayed only a few glycan ions as the result of low sample concentration or the presence of contamination. Ion mobility also enabled uncontaminated CID spectra to be obtained from glycans when their molecular ions displayed coincidence with ions from fragments or multiply charged ions with similar m/z values. This technique proved to be invaluable for removing extraneous ions from many CID spectra. The presence of such ions often produces spectra that are difficult to interpret. Most CID spectra, even those from abundant glycan constituents, benefited from such clean-up, showing that the extra dimension provided by ion mobility was invaluable for studies of this type.
authors.affiliation: Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK. david.harvey@bioch.ox.ac.uk.|Department of Biological Sciences, University of Warwick, Coventry, CV47AL, UK. david.harvey@bioch.ox.ac.uk.
authors.forename: David J
authors.initials: DJ
authors.lastname: Harvey
authors.affiliation: Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors.forename: Max
authors.initials: M
authors.lastname: Crispin
authors.affiliation: Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors.forename: Camille
authors.initials: C
authors.lastname: Bonomelli
authors.affiliation: Department of Biological Sciences, University of Warwick, Coventry, CV47AL, UK.
authors.forename: Jim H
authors.initials: JH
authors.lastname: Scrivens
mesh_terms: D002236:Carbohydrate Conformation; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D007477:Ions / Q000032:analysis / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards; D053719:Tandem Mass Spectrometry; D014764:Viral Proteins / Q000032:analysis* / Q000737:chemistry
chemical_list: D006023:Glycoproteins; D007477:Ions; D011134:Polysaccharides; D014764:Viral Proteins
keywords: CID; Contamination; Ion mobility; N-glycans