Tandem mass spectrometry of isomeric aniline‐labeled N‐glycans separated on porous graphitic carbon: Revealing the attachment position of terminal sialic acids and structures of neutral glycans
Identifieur interne : 000610 ( Main/Exploration ); précédent : 000609; suivant : 000611Tandem mass spectrometry of isomeric aniline‐labeled N‐glycans separated on porous graphitic carbon: Revealing the attachment position of terminal sialic acids and structures of neutral glycans
Auteurs : Claudia Michael [Autriche] ; Andreas M. Rizzi [Autriche]Source :
- Rapid Communications in Mass Spectrometry [ 0951-4198 ] ; 2015-07-15.
Abstract
Rationale: Quantitative monitoring of changes in the N‐glycome upon disease has gained significance in the context of biomarker discovery. Separation and quantification of isobaric glycan isomers can be attained by using high‐performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI‐MS). Collision‐induced dissociation (CID)‐based fragmentation of separated isobaric glycans is evaluated in respect to its potential of providing fragment ions specific for the linkage positions of terminal sialic acids and the presence of intersecting GlcNAc moieties, respectively. Methods: N‐Glycans were labeled via reductive amination using 12C6‐aniline and 13C6‐aniline as isotope‐coded labeling reagents. The differently labeled glycans were merged and separated into various species using a porous graphitic carbon (PGC) stationary phase. Identification of structural features of separated isobaric isomers was performed by CID‐based tandem mass spectrometry (MS/MS) carried out in a quadrupole time‐of‐flight (QqTOF) or a quadrupole ion‐trap (IT) mass spectrometer. Results: Working in the negative ion mode, new diagnostic CID fragment ions could be found that are indicative for the α2,6‐type linkage of sialic acids. Other diagnostic ions, identified before as being indicative for the substitution of the 6‐antenna, could be confirmed as being of relevance also in the case of aniline labeling. In the positive ion mode, CID fragment ions indicative for the structure of short neutral N‐glycans were identified. Conclusions: One new diagnostic ion specific for the linkage position of the terminal sialic acids and one for the presence of bisecting GlcNAc in N‐glycans were identified. The aniline label introduced for improved relative quantitation in MS1 was found not to significantly alter the CID fragmentation patterns that were reported previously by other authors for unlabeled/reduced glycans or for glycans with more polar labels. Copyright © 2015 John Wiley & Sons, Ltd.
Url:
DOI: 10.1002/rcm.7208
Affiliations:
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<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Tandem mass spectrometry of isomeric aniline‐labeled N‐glycans separated on porous graphitic carbon: Revealing the attachment position of terminal sialic acids and structures of neutral glycans</title><author><name sortKey="Michael, Claudia" sort="Michael, Claudia" uniqKey="Michael C" first="Claudia" last="Michael">Claudia Michael</name></author><author><name sortKey="Rizzi, Andreas M" sort="Rizzi, Andreas M" uniqKey="Rizzi A" first="Andreas M." last="Rizzi">Andreas M. 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Rizzi</name><affiliation wicri:level="3"><country xml:lang="fr">Autriche</country><wicri:regionArea>Institute of Analytical Chemistry, University of Vienna, Währinger Straße 38, 1090, Vienna</wicri:regionArea><placeName><settlement type="city">Vienne (Autriche)</settlement><region nuts="2" type="province">Vienne (Autriche)</region></placeName></affiliation><affiliation></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Autriche</country></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Rapid Communications in Mass Spectrometry</title><title level="j" type="alt">RAPID COMMUNICATIONS IN MASS SPECTROMETRY</title><idno type="ISSN">0951-4198</idno><idno type="eISSN">1097-0231</idno><imprint><biblScope unit="vol">29</biblScope><biblScope unit="issue">13</biblScope><biblScope unit="page" from="1268">1268</biblScope><biblScope unit="page" to="1278">1278</biblScope><biblScope unit="page-count">11</biblScope><date type="published" when="2015-07-15">2015-07-15</date></imprint><idno type="ISSN">0951-4198</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0951-4198</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract">Rationale: Quantitative monitoring of changes in the N‐glycome upon disease has gained significance in the context of biomarker discovery. Separation and quantification of isobaric glycan isomers can be attained by using high‐performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI‐MS). Collision‐induced dissociation (CID)‐based fragmentation of separated isobaric glycans is evaluated in respect to its potential of providing fragment ions specific for the linkage positions of terminal sialic acids and the presence of intersecting GlcNAc moieties, respectively. Methods: N‐Glycans were labeled via reductive amination using 12C6‐aniline and 13C6‐aniline as isotope‐coded labeling reagents. The differently labeled glycans were merged and separated into various species using a porous graphitic carbon (PGC) stationary phase. Identification of structural features of separated isobaric isomers was performed by CID‐based tandem mass spectrometry (MS/MS) carried out in a quadrupole time‐of‐flight (QqTOF) or a quadrupole ion‐trap (IT) mass spectrometer. Results: Working in the negative ion mode, new diagnostic CID fragment ions could be found that are indicative for the α2,6‐type linkage of sialic acids. Other diagnostic ions, identified before as being indicative for the substitution of the 6‐antenna, could be confirmed as being of relevance also in the case of aniline labeling. In the positive ion mode, CID fragment ions indicative for the structure of short neutral N‐glycans were identified. Conclusions: One new diagnostic ion specific for the linkage position of the terminal sialic acids and one for the presence of bisecting GlcNAc in N‐glycans were identified. The aniline label introduced for improved relative quantitation in MS1 was found not to significantly alter the CID fragmentation patterns that were reported previously by other authors for unlabeled/reduced glycans or for glycans with more polar labels. Copyright © 2015 John Wiley & Sons, Ltd.</div></front></TEI><affiliations><list><country><li>Autriche</li></country><region><li>Vienne (Autriche)</li></region><settlement><li>Vienne (Autriche)</li></settlement></list><tree><country name="Autriche"><region name="Vienne (Autriche)"><name sortKey="Michael, Claudia" sort="Michael, Claudia" uniqKey="Michael C" first="Claudia" last="Michael">Claudia Michael</name></region><name sortKey="Rizzi, Andreas M" sort="Rizzi, Andreas M" uniqKey="Rizzi A" first="Andreas M." last="Rizzi">Andreas M. Rizzi</name><name sortKey="Rizzi, Andreas M" sort="Rizzi, Andreas M" uniqKey="Rizzi A" first="Andreas M." last="Rizzi">Andreas M. Rizzi</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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