Characterization of VSV-GP morphology by cryo-EM imaging and SEC-MALS

Vesicular stomatitis virus expressing the glycoprotein of the lymphocytic choriomeningitis virus (VSV-GP) is a promising platform for oncolytic viruses and cancer vaccines. In this work, cryoelectron microscopy (cryo-EM) imaging was employed to directly visualize VSV-GP particles. Several different...

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Main Authors: Dongyue Xin, Leela Kurien, Katherine Briggs, Adrian Schimek, Richard Dambra, Daniel Hochdorfer, Tanja A. Arnouk, Marija Brgles, Saurabh Gautam, Dominik Hotter, Johannes Solzin, Thomas Kriehuber, Joseph Ashour, Adam Vigil, Michael Hawley, Xiaorong He
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:Molecular Therapy: Methods & Clinical Development
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Online Access:http://www.sciencedirect.com/science/article/pii/S2329050125000245
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author Dongyue Xin
Leela Kurien
Katherine Briggs
Adrian Schimek
Richard Dambra
Daniel Hochdorfer
Tanja A. Arnouk
Marija Brgles
Saurabh Gautam
Dominik Hotter
Johannes Solzin
Thomas Kriehuber
Joseph Ashour
Adam Vigil
Michael Hawley
Xiaorong He
author_facet Dongyue Xin
Leela Kurien
Katherine Briggs
Adrian Schimek
Richard Dambra
Daniel Hochdorfer
Tanja A. Arnouk
Marija Brgles
Saurabh Gautam
Dominik Hotter
Johannes Solzin
Thomas Kriehuber
Joseph Ashour
Adam Vigil
Michael Hawley
Xiaorong He
author_sort Dongyue Xin
collection DOAJ
description Vesicular stomatitis virus expressing the glycoprotein of the lymphocytic choriomeningitis virus (VSV-GP) is a promising platform for oncolytic viruses and cancer vaccines. In this work, cryoelectron microscopy (cryo-EM) imaging was employed to directly visualize VSV-GP particles. Several different subpopulations of virus particle morphology were observed. Definition and fraction counting of subpopulations enabled quantitative comparison of subpopulation profiles between several VSV-GP samples. In developing an orthogonal method with higher throughput, we showed that the morphological profile of the VSV-GP particles can be characterized by size exclusion chromatography coupled with a multi-angle light scattering detector (SEC-MALS) based on a novel shape-based separation mechanism. Together, the two complementary techniques enable the analysis of morphological profile for VSV-GP and potentially other non-spherical viruses or nanoparticles.
format Article
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institution DOAJ
issn 2329-0501
language English
publishDate 2025-03-01
publisher Elsevier
record_format Article
series Molecular Therapy: Methods & Clinical Development
spelling doaj-art-e3f041d35bf9495aa01e9af844929fc42025-08-20T03:00:31ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012025-03-0133110142910.1016/j.omtm.2025.101429Characterization of VSV-GP morphology by cryo-EM imaging and SEC-MALSDongyue Xin0Leela Kurien1Katherine Briggs2Adrian Schimek3Richard Dambra4Daniel Hochdorfer5Tanja A. Arnouk6Marija Brgles7Saurabh Gautam8Dominik Hotter9Johannes Solzin10Thomas Kriehuber11Joseph Ashour12Adam Vigil13Michael Hawley14Xiaorong He15Material and Analytical Sciences, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USA; Corresponding author: Dongyue Xin, Material and Analytical Sciences, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USA.Material and Analytical Sciences, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USAMaterial and Analytical Sciences, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USAViraTherapeutics GmbH, Innsbruck, AustriaDrug Metabolism and Pharmacokinetics, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USAViral Therapeutics Center, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyViral Therapeutics Center, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyViral Therapeutics Center, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyViraTherapeutics GmbH, Innsbruck, Austria; Boehringer Ingelheim International GmbH, Ingelheim, GermanyViral Therapeutics Center, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyViral Therapeutics Center, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyViral Therapeutics Center, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, GermanyDrug Metabolism and Pharmacokinetics, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USADrug Metabolism and Pharmacokinetics, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USAMaterial and Analytical Sciences, Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USABoehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT, USAVesicular stomatitis virus expressing the glycoprotein of the lymphocytic choriomeningitis virus (VSV-GP) is a promising platform for oncolytic viruses and cancer vaccines. In this work, cryoelectron microscopy (cryo-EM) imaging was employed to directly visualize VSV-GP particles. Several different subpopulations of virus particle morphology were observed. Definition and fraction counting of subpopulations enabled quantitative comparison of subpopulation profiles between several VSV-GP samples. In developing an orthogonal method with higher throughput, we showed that the morphological profile of the VSV-GP particles can be characterized by size exclusion chromatography coupled with a multi-angle light scattering detector (SEC-MALS) based on a novel shape-based separation mechanism. Together, the two complementary techniques enable the analysis of morphological profile for VSV-GP and potentially other non-spherical viruses or nanoparticles.http://www.sciencedirect.com/science/article/pii/S2329050125000245vesicular stomatitis virusVSV-GPvirus particle morphologycryo-EMSEC-MALSanalytical HPLC
spellingShingle Dongyue Xin
Leela Kurien
Katherine Briggs
Adrian Schimek
Richard Dambra
Daniel Hochdorfer
Tanja A. Arnouk
Marija Brgles
Saurabh Gautam
Dominik Hotter
Johannes Solzin
Thomas Kriehuber
Joseph Ashour
Adam Vigil
Michael Hawley
Xiaorong He
Characterization of VSV-GP morphology by cryo-EM imaging and SEC-MALS
Molecular Therapy: Methods & Clinical Development
vesicular stomatitis virus
VSV-GP
virus particle morphology
cryo-EM
SEC-MALS
analytical HPLC
title Characterization of VSV-GP morphology by cryo-EM imaging and SEC-MALS
title_full Characterization of VSV-GP morphology by cryo-EM imaging and SEC-MALS
title_fullStr Characterization of VSV-GP morphology by cryo-EM imaging and SEC-MALS
title_full_unstemmed Characterization of VSV-GP morphology by cryo-EM imaging and SEC-MALS
title_short Characterization of VSV-GP morphology by cryo-EM imaging and SEC-MALS
title_sort characterization of vsv gp morphology by cryo em imaging and sec mals
topic vesicular stomatitis virus
VSV-GP
virus particle morphology
cryo-EM
SEC-MALS
analytical HPLC
url http://www.sciencedirect.com/science/article/pii/S2329050125000245
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