Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans

David J. Harvey, Charlotte A. Scarff, Matthew Edgeworth, Weston B. Struwe, Kevin Pagel, Konstantinos Thalassinos, Max Crispin, James Scrivens

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The isomeric structure of high-mannose N-glycans can significantly impact biological recognition events. Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. In some cases, asymmetric ATDs were observed, but no isomers could be detected by fragmentation. In these cases, it was assumed that conformers or anomers were being separated. Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed.
Original languageEnglish
Pages (from-to)219-235
JournalJournal of Mass Spectrometry
Volume51
Issue number3
DOIs
Publication statusPublished - 4 Mar 2016

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Mannose
Isomers
Polysaccharides
Negative ions
Ions
Mass spectrometry
Thyroglobulin
Ovalbumin
Dextrans
Viruses
Ionization
Glycoproteins
Positive ions

Cite this

Harvey, D. J., Scarff, C. A., Edgeworth, M., Struwe, W. B., Pagel, K., Thalassinos, K., ... Scrivens, J. (2016). Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. Journal of Mass Spectrometry, 51(3), 219-235. https://doi.org/10.1002/jms.3738
Harvey, David J. ; Scarff, Charlotte A. ; Edgeworth, Matthew ; Struwe, Weston B. ; Pagel, Kevin ; Thalassinos, Konstantinos ; Crispin, Max ; Scrivens, James. / Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. In: Journal of Mass Spectrometry. 2016 ; Vol. 51, No. 3. pp. 219-235.
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Harvey, DJ, Scarff, CA, Edgeworth, M, Struwe, WB, Pagel, K, Thalassinos, K, Crispin, M & Scrivens, J 2016, 'Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans', Journal of Mass Spectrometry, vol. 51, no. 3, pp. 219-235. https://doi.org/10.1002/jms.3738

Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. / Harvey, David J.; Scarff, Charlotte A.; Edgeworth, Matthew; Struwe, Weston B.; Pagel, Kevin; Thalassinos, Konstantinos; Crispin, Max; Scrivens, James.

In: Journal of Mass Spectrometry, Vol. 51, No. 3, 04.03.2016, p. 219-235.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Harvey, David J.

AU - Scarff, Charlotte A.

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AU - Thalassinos, Konstantinos

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AB - The isomeric structure of high-mannose N-glycans can significantly impact biological recognition events. Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. In some cases, asymmetric ATDs were observed, but no isomers could be detected by fragmentation. In these cases, it was assumed that conformers or anomers were being separated. Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed.

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Harvey DJ, Scarff CA, Edgeworth M, Struwe WB, Pagel K, Thalassinos K et al. Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. Journal of Mass Spectrometry. 2016 Mar 4;51(3):219-235. https://doi.org/10.1002/jms.3738