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...
Guardado en:
Autores principales: | , , , , , , , , |
---|---|
Formato: | article |
Lenguaje: | EN |
Publicado: |
Elsevier
2021
|
Materias: | |
Acceso en línea: | https://doaj.org/article/80ee3b897ec94e55bc1c335e48d70703 |
Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
id |
oai:doaj.org-article:80ee3b897ec94e55bc1c335e48d70703 |
---|---|
record_format |
dspace |
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 |
work_keys_str_mv |
AT jacobpvanvloten productionandpurificationofhightiterorfvforpreclinicalstudiesinvaccinologyandcancertherapy AT jessicaaminott productionandpurificationofhightiterorfvforpreclinicalstudiesinvaccinologyandcancertherapy AT thomasmmcausland productionandpurificationofhightiterorfvforpreclinicalstudiesinvaccinologyandcancertherapy AT joellecingrao productionandpurificationofhightiterorfvforpreclinicalstudiesinvaccinologyandcancertherapy AT lisaasantry productionandpurificationofhightiterorfvforpreclinicalstudiesinvaccinologyandcancertherapy AT grantmcfadden productionandpurificationofhightiterorfvforpreclinicalstudiesinvaccinologyandcancertherapy AT jamesjpetrik productionandpurificationofhightiterorfvforpreclinicalstudiesinvaccinologyandcancertherapy AT byramwbridle productionandpurificationofhightiterorfvforpreclinicalstudiesinvaccinologyandcancertherapy AT sarahkwootton productionandpurificationofhightiterorfvforpreclinicalstudiesinvaccinologyandcancertherapy |
_version_ |
1718445283666296832 |