<div class="csl-bib-body">
<div class="csl-entry">Zoratto, S., Weiss, V., van der Horst, J., Commandeur, J., Buengener, C., Foettinger-Vacha, A., Pletzenauer, R., Graninger, M., & Allmaier, G. (2021). Molecular weight determination of adeno-associate virus serotype 8 virus-like particle either carrying or lacking genome via native nES gas-phase electrophoretic molecular mobility analysis and nESI QRTOF mass spectrometry. <i>Journal of Mass Spectrometry</i>, <i>56</i>(11), Article e4786. https://doi.org/10.1002/jms.4786</div>
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dc.identifier.issn
1076-5174
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dc.identifier.uri
http://hdl.handle.net/20.500.12708/138767
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dc.description.abstract
Virus-like particles (VLPs) are proteinaceous shells derived from viruses lacking any viral genomic material. Adeno-associated virus (AAV) is a non-enveloped icosahedral virus used as VLP delivery system in gene therapy (GT). Its success as vehicle for GT is due to its selective tropism, high level of transduction, and low immunogenicity. In this study, two preparations of AAV serotype 8 (AAV8) VLPs either carrying or lacking completely genomic cargo (i.e., non-viral ssDNA) have been investigated by means of a native nano-electrospray gas-phase electrophoretic mobility molecular analyzer (GEMMA) (native nES GEMMA) and native nano-electrospray ionization quadrupole reflectron time-of-flight mass spectrometry (MS) (native nESI QRTOF MS). nES GEMMA is based on electrophoretic mobility principles: single-charge nanoparticles (NPs), that is, AAV8 particle, are separated in a laminar sheath flow of dry, particle-free air and a tunable orthogonal electric field. Thus, the electrophoretic mobility diameter (EMD) of a bio-NP (i.e., diameter of globular nano-objects) is obtained at atmospheric pressure, which can be converted into its MW based on a correlation. First is the native nESI QRTOF. MS's goal is to keep the native biological conformation of an analyte during the passage into the vacuum. Subsequently, highly accurate MW values are obtained from multiple-charged species after deconvolution. However, once applied to the analysis of megadalton species, native MS is challenging and requires customized instrumental modifications not readily available on standard devices. Hence, the analysis of AAV8 VLPs via native MS in our hands did not produce a defined charge state assignment, that is, charge deconvolution for exact MW determination was not possible. Nonetheless, the method we present is capable to estimate the MW of VLPs by combining the results from native nES GEMMA and native ESI QRTOF MS. In detail, our findings show a MW of 3.7 and 5.0 MDa for AAV8 VLPs either lacking or carrying an engineered genome, respectively.
en
dc.language.iso
en
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dc.publisher
WILEY
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dc.relation.ispartof
Journal of Mass Spectrometry
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dc.subject
Spectroscopy
en
dc.subject
adeno-associated virus
en
dc.subject
AAV
en
dc.subject
GEMMA
en
dc.subject
gene therapy platform
en
dc.subject
native MS
en
dc.title
Molecular weight determination of adeno-associate virus serotype 8 virus-like particle either carrying or lacking genome via native nES gas-phase electrophoretic molecular mobility analysis and nESI QRTOF mass spectrometry
en
dc.type
Artikel
de
dc.type
Article
en
dc.type.category
Original Research Article
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tuw.container.volume
56
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tuw.container.issue
11
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tuw.journal.peerreviewed
true
-
tuw.peerreviewed
true
-
wb.publication.intCoWork
International Co-publication
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tuw.researchTopic.id
M6
-
tuw.researchTopic.name
Biological and Bioactive Materials
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tuw.researchTopic.value
100
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dcterms.isPartOf.title
Journal of Mass Spectrometry
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tuw.publication.orgunit
E164-01-1 - Forschungsgruppe Massenspektrometrische Bio- und Polymeranalytik
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tuw.publisher.doi
10.1002/jms.4786
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dc.identifier.articleid
e4786
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dc.identifier.eissn
1096-9888
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dc.description.numberOfPages
8
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tuw.author.orcid
0000-0002-1491-3404
-
tuw.author.orcid
0000-0002-0056-6819
-
tuw.author.orcid
0000-0002-1438-9462
-
wb.sci
true
-
wb.sciencebranch
Chemie
-
wb.sciencebranch
Biologie
-
wb.sciencebranch.oefos
1040
-
wb.sciencebranch.oefos
1060
-
wb.facultyfocus
Bioscience Technology
de
wb.facultyfocus
Bioscience Technology
en
wb.facultyfocus.faculty
E150
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item.openairetype
research article
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item.fulltext
no Fulltext
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item.openairecristype
http://purl.org/coar/resource_type/c_2df8fbb1
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item.grantfulltext
restricted
-
item.cerifentitytype
Publications
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item.languageiso639-1
en
-
crisitem.author.dept
E164-01-1 - Forschungsgruppe Massenspektrometrische Bio- und Polymeranalytik
-
crisitem.author.dept
E164-01-1 - Forschungsgruppe Massenspektrometrische Bio- und Polymeranalytik
-
crisitem.author.dept
E164-01-1 - Forschungsgruppe Massenspektrometrische Bio- und Polymeranalytik
-
crisitem.author.orcid
0000-0002-1491-3404
-
crisitem.author.orcid
0000-0002-0056-6819
-
crisitem.author.orcid
0000-0002-1438-9462
-
crisitem.author.parentorg
E164-01 - Forschungsbereich Imaging und Instrumentelle Analytische Chemie
-
crisitem.author.parentorg
E164-01 - Forschungsbereich Imaging und Instrumentelle Analytische Chemie
-
crisitem.author.parentorg
E164-01 - Forschungsbereich Imaging und Instrumentelle Analytische Chemie