<div class="csl-bib-body">
<div class="csl-entry">Soher, E. (2016). <i>Method development for quantifying protein modifications using liquid chromatography hyphenated to tandem mass spectrometry</i> [Diploma Thesis, Technische Universität Wien]. reposiTUm. https://doi.org/10.34726/hss.2016.24554</div>
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dc.identifier.uri
https://doi.org/10.34726/hss.2016.24554
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dc.identifier.uri
http://hdl.handle.net/20.500.12708/6568
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dc.description
Zusammenfassung in deutscher Sprache
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dc.description
Abweichender Titel nach Übersetzung der Verfasserin/des Verfassers
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dc.description.abstract
Zearalenone (ZEN) is a mycotoxin, which is produced by various species of the fungus Fusarium, such as F. graminearum, F. equiseti and F.culmorum. Since this fungus is common to grow on crops, food and feed are often contaminated with the mycotoxin. Romer Labs developed a quick test, in form of a lateral flow device (LFD), to determine the amount of contamination on grain. An important constituent for this LFD is zearalenone - carboxymethyl oxime (ZEN-CMO) coupled to conalbumin (CON). Since it is always desirable to produce at the lowest possible cost, it is of interest whether different synthesis strategies can lead to the same results in terms of modification efficiency on the protein, while reducing various synthesis steps or chemicals in use. Therefore the goal was to establish a liquid chromatography - tandem mass spectrometry method (LC-MS/MS) to quantify ZEN-CMO modifications introduced onto the target protein. For detailed protein characterization, the proteins were digested and analyzed on a nano-LC-electrospray ionization (ESI)-ion trap mass spectrometer performing collision induced dissociation (CID) fragmentation for peptide identification and modification localization. In order to maximize protein sequence coverage, two proteases were tested. Immobilized trypsin was compared to a trypsin/LysC mix. Further method development for quantification was performed on an ultra high performance liquid chromatography (UPLC) - ESI - triple quadrupole instrument. We found trypsin/LysC to perform best in terms of sequence coverage after in solution digestion and peptide desalting. It was possible to identify 27 out of 59 possible Lysine modification sites and it was found that most of the identified protein modifications were located on the protein-s surface. Furthermore, fragmentation mass spectra gave insight into fragmentation mechanisms of ZEN-CMO and ZEN-CMO modified peptides using an ion trap mass analyzer, exhibiting reporter fragment ions which were further used to develop a quantification method applying triple quadrupole MS technology.
en
dc.language
English
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dc.language.iso
en
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dc.rights.uri
http://rightsstatements.org/vocab/InC/1.0/
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dc.subject
Massenspektrometrie
de
dc.subject
Proteinkonjugate
de
dc.subject
Mass Spectrometry
en
dc.subject
Protein Conjugates
en
dc.title
Method development for quantifying protein modifications using liquid chromatography hyphenated to tandem mass spectrometry
en
dc.title.alternative
Etablierung einer LC-MS/MS Methode zur Quantitativen Bestimmung von Peptidmodifikationen
de
dc.type
Thesis
en
dc.type
Hochschulschrift
de
dc.rights.license
In Copyright
en
dc.rights.license
Urheberrechtsschutz
de
dc.identifier.doi
10.34726/hss.2016.24554
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dc.contributor.affiliation
TU Wien, Österreich
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dc.rights.holder
Elisabeth Soher
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dc.publisher.place
Wien
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tuw.version
vor
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tuw.thesisinformation
Technische Universität Wien
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tuw.publication.orgunit
E164 - Institut für Chemische Technologien und Analytik
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dc.type.qualificationlevel
Diploma
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dc.identifier.libraryid
AC13093715
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dc.description.numberOfPages
167
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dc.identifier.urn
urn:nbn:at:at-ubtuw:1-1555
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dc.thesistype
Diplomarbeit
de
dc.thesistype
Diploma Thesis
en
dc.rights.identifier
In Copyright
en
dc.rights.identifier
Urheberrechtsschutz
de
tuw.advisor.staffStatus
staff
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tuw.advisor.orcid
0000-0002-8060-7851
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item.fulltext
with Fulltext
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item.cerifentitytype
Publications
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item.mimetype
application/pdf
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item.openairecristype
http://purl.org/coar/resource_type/c_bdcc
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item.languageiso639-1
en
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item.openaccessfulltext
Open Access
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item.openairetype
master thesis
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item.grantfulltext
open
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crisitem.author.dept
E164 - Institut für Chemische Technologien und Analytik