Ion mobility mass spectrometry for ion recovery and clean-up of MS and MS/MS spectra obtained from low abundance viral samples
Ion mobility mass spectrometry for ion recovery and clean-up of MS and MS/MS spectra obtained from low abundance viral samples
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.
CID, Contamination, Ion mobility, N-glycans
1754-1767
Harvey, David J.
8bb24417-3852-4b1f-827b-0d5d2c176744
Crispin, Max
cd980957-0943-4b89-b2b2-710f01f33bc9
Bonomelli, Camille
51edb32c-85d0-45be-b050-b075cc3f6c28
Scrivens, Jim H.
12019574-e300-490b-8a96-50fa927bfd89
15 October 2015
Harvey, David J.
8bb24417-3852-4b1f-827b-0d5d2c176744
Crispin, Max
cd980957-0943-4b89-b2b2-710f01f33bc9
Bonomelli, Camille
51edb32c-85d0-45be-b050-b075cc3f6c28
Scrivens, Jim H.
12019574-e300-490b-8a96-50fa927bfd89
Harvey, David J., Crispin, Max, Bonomelli, Camille and Scrivens, Jim H.
(2015)
Ion mobility mass spectrometry for ion recovery and clean-up of MS and MS/MS spectra obtained from low abundance viral samples.
Journal of the American Society for Mass Spectrometry, 26 (10), .
(doi:10.1007/s13361-015-1163-5).
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.
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More information
e-pub ahead of print date: 24 July 2015
Published date: 15 October 2015
Keywords:
CID, Contamination, Ion mobility, N-glycans
Identifiers
Local EPrints ID: 414317
URI: http://eprints.soton.ac.uk/id/eprint/414317
ISSN: 1044-0305
PURE UUID: a41eeb25-938a-4b45-8b00-30a9a0fcb02e
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Date deposited: 26 Sep 2017 16:30
Last modified: 16 Mar 2024 04:30
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Contributors
Author:
David J. Harvey
Author:
Camille Bonomelli
Author:
Jim H. Scrivens
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