An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells

Measles virus envelope pseudotyped LV (MV-LV) can achieve high B cell transduction rates (up to 50%), but suffers from low titers. To overcome current limitations, we developed an optimized MV-LV production protocol that achieved consistent B cell transduction efficiency up to 75%. We detail this pr...

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Detalles Bibliográficos
Autores principales: Vamva, Eirini, Ozog, Stosh, Verhoeyen, Els, James, Richard G., Rawlings, David J., Torbett, Bruce E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8914380/
https://www.ncbi.nlm.nih.gov/pubmed/35284833
http://dx.doi.org/10.1016/j.xpro.2022.101228
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author Vamva, Eirini
Ozog, Stosh
Verhoeyen, Els
James, Richard G.
Rawlings, David J.
Torbett, Bruce E.
author_facet Vamva, Eirini
Ozog, Stosh
Verhoeyen, Els
James, Richard G.
Rawlings, David J.
Torbett, Bruce E.
author_sort Vamva, Eirini
collection PubMed
description Measles virus envelope pseudotyped LV (MV-LV) can achieve high B cell transduction rates (up to 50%), but suffers from low titers. To overcome current limitations, we developed an optimized MV-LV production protocol that achieved consistent B cell transduction efficiency up to 75%. We detail this protocol along with analytical assays to assess the results of MV-LV mediated B cell transduction, including flow cytometry for B cell phenotypic characterization and measurement of transduction efficiency, and ddPCR for VCN analysis.
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spelling pubmed-89143802022-03-12 An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells Vamva, Eirini Ozog, Stosh Verhoeyen, Els James, Richard G. Rawlings, David J. Torbett, Bruce E. STAR Protoc Protocol Measles virus envelope pseudotyped LV (MV-LV) can achieve high B cell transduction rates (up to 50%), but suffers from low titers. To overcome current limitations, we developed an optimized MV-LV production protocol that achieved consistent B cell transduction efficiency up to 75%. We detail this protocol along with analytical assays to assess the results of MV-LV mediated B cell transduction, including flow cytometry for B cell phenotypic characterization and measurement of transduction efficiency, and ddPCR for VCN analysis. Elsevier 2022-03-08 /pmc/articles/PMC8914380/ /pubmed/35284833 http://dx.doi.org/10.1016/j.xpro.2022.101228 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Vamva, Eirini
Ozog, Stosh
Verhoeyen, Els
James, Richard G.
Rawlings, David J.
Torbett, Bruce E.
An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells
title An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells
title_full An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells
title_fullStr An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells
title_full_unstemmed An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells
title_short An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells
title_sort optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary b cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8914380/
https://www.ncbi.nlm.nih.gov/pubmed/35284833
http://dx.doi.org/10.1016/j.xpro.2022.101228
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