Production and purification of high-titer OrfV for preclinical studies in vaccinology and cancer therapy

Poxviruses have been used extensively as vaccine vectors for human and veterinary medicine and have recently entered the clinical realm as immunotherapies for cancer. We present a comprehensive method for producing high-quality lots of the poxvirus Parapoxvirus ovis (OrfV) for use in preclinical mod...

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Autores principales: Jacob P. van Vloten, Jessica A. Minott, Thomas M. McAusland, Joelle C. Ingrao, Lisa A. Santry, Grant McFadden, James J. Petrik, Byram W. Bridle, Sarah K. Wootton
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Publicado: Elsevier 2021
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Acceso en línea:https://doaj.org/article/80ee3b897ec94e55bc1c335e48d70703
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spelling oai:doaj.org-article:80ee3b897ec94e55bc1c335e48d707032021-11-04T04:32:14ZProduction and purification of high-titer OrfV for preclinical studies in vaccinology and cancer therapy2329-050110.1016/j.omtm.2021.08.004https://doaj.org/article/80ee3b897ec94e55bc1c335e48d707032021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2329050121001303https://doaj.org/toc/2329-0501Poxviruses have been used extensively as vaccine vectors for human and veterinary medicine and have recently entered the clinical realm as immunotherapies for cancer. We present a comprehensive method for producing high-quality lots of the poxvirus Parapoxvirus ovis (OrfV) for use in preclinical models of vaccinology and cancer therapy. OrfV is produced using a permissive sheep skin-derived cell line and is released from infected cells by repeated freeze-thaw combined with sonication. We present two methods for isolation and purification of bulk virus. Isolated virus is concentrated to high titer using polyethylene glycol to produce the final in vivo-grade product. We also describe methods for quantifying OrfV infectious virions and determining genomic copy number to evaluate virus stocks. The methods herein will provide researchers with the ability to produce high-quality, high-titer OrfV for use in preclinical studies, and support the translation of OrfV-derived technologies into the clinic.Jacob P. van VlotenJessica A. MinottThomas M. McAuslandJoelle C. IngraoLisa A. SantryGrant McFaddenJames J. PetrikByram W. BridleSarah K. WoottonElsevierarticleParapoxvirus ovisOrfVtangential flow filtrationvirus purificationvaccinologycancer therapyGeneticsQH426-470CytologyQH573-671ENMolecular Therapy: Methods & Clinical Development, Vol 23, Iss , Pp 434-447 (2021)
institution DOAJ
collection DOAJ
language EN
topic Parapoxvirus ovis
OrfV
tangential flow filtration
virus purification
vaccinology
cancer therapy
Genetics
QH426-470
Cytology
QH573-671
spellingShingle Parapoxvirus ovis
OrfV
tangential flow filtration
virus purification
vaccinology
cancer therapy
Genetics
QH426-470
Cytology
QH573-671
Jacob P. van Vloten
Jessica A. Minott
Thomas M. McAusland
Joelle C. Ingrao
Lisa A. Santry
Grant McFadden
James J. Petrik
Byram W. Bridle
Sarah K. Wootton
Production and purification of high-titer OrfV for preclinical studies in vaccinology and cancer therapy
description Poxviruses have been used extensively as vaccine vectors for human and veterinary medicine and have recently entered the clinical realm as immunotherapies for cancer. We present a comprehensive method for producing high-quality lots of the poxvirus Parapoxvirus ovis (OrfV) for use in preclinical models of vaccinology and cancer therapy. OrfV is produced using a permissive sheep skin-derived cell line and is released from infected cells by repeated freeze-thaw combined with sonication. We present two methods for isolation and purification of bulk virus. Isolated virus is concentrated to high titer using polyethylene glycol to produce the final in vivo-grade product. We also describe methods for quantifying OrfV infectious virions and determining genomic copy number to evaluate virus stocks. The methods herein will provide researchers with the ability to produce high-quality, high-titer OrfV for use in preclinical studies, and support the translation of OrfV-derived technologies into the clinic.
format article
author Jacob P. van Vloten
Jessica A. Minott
Thomas M. McAusland
Joelle C. Ingrao
Lisa A. Santry
Grant McFadden
James J. Petrik
Byram W. Bridle
Sarah K. Wootton
author_facet Jacob P. van Vloten
Jessica A. Minott
Thomas M. McAusland
Joelle C. Ingrao
Lisa A. Santry
Grant McFadden
James J. Petrik
Byram W. Bridle
Sarah K. Wootton
author_sort Jacob P. van Vloten
title Production and purification of high-titer OrfV for preclinical studies in vaccinology and cancer therapy
title_short Production and purification of high-titer OrfV for preclinical studies in vaccinology and cancer therapy
title_full Production and purification of high-titer OrfV for preclinical studies in vaccinology and cancer therapy
title_fullStr Production and purification of high-titer OrfV for preclinical studies in vaccinology and cancer therapy
title_full_unstemmed Production and purification of high-titer OrfV for preclinical studies in vaccinology and cancer therapy
title_sort production and purification of high-titer orfv for preclinical studies in vaccinology and cancer therapy
publisher Elsevier
publishDate 2021
url https://doaj.org/article/80ee3b897ec94e55bc1c335e48d70703
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