Reference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells

Abstract Lentiviral vectors (LV) have emerged as a robust technology for therapeutic gene delivery into human cells as advanced medicinal products. As these products are increasingly commercialized, there are concomitant demands for their characterization to ensure safety, efficacy and consistency....

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Autores principales:	Barbara S. Paugh, Lajos Baranyi, Andre Roy, Hua-Jun He, Lindsay Harris, Kenneth D. Cole, Moria Artlip, Caroline Raimund, Patricia S. Langan, Srikanta Jana, Rimas J. Orentas, Sheng Lin-Gibson, Winfried Krueger, Boro Dropulić
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3dd0bd30801246de9358d8be2c031336
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spelling	oai:doaj.org-article:3dd0bd30801246de9358d8be2c0313362021-12-02T15:23:10ZReference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells10.1038/s41598-020-79698-w2045-2322https://doaj.org/article/3dd0bd30801246de9358d8be2c0313362021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-79698-whttps://doaj.org/toc/2045-2322Abstract Lentiviral vectors (LV) have emerged as a robust technology for therapeutic gene delivery into human cells as advanced medicinal products. As these products are increasingly commercialized, there are concomitant demands for their characterization to ensure safety, efficacy and consistency. Standards are essential for accurately measuring parameters for such product characterization. A critical parameter is the vector copy number (VCN) which measures the genetic dose of a transgene present in gene-modified cells. Here we describe a set of clonal Jurkat cell lines with defined copy numbers of a reference lentiviral vector integrated into their genomes. Genomic DNA was characterized for copy number, genomic integrity and integration coordinates and showed uniform performance across independent quantitative PCR assays. Stability studies during continuous long-term culture demonstrated sustained renewability of the reference standard source material. DNA from the Jurkat VCN standards would be useful for control of quantitative PCR assays for VCN determination in LV gene-modified cellular products and clinical samples.Barbara S. PaughLajos BaranyiAndre RoyHua-Jun HeLindsay HarrisKenneth D. ColeMoria ArtlipCaroline RaimundPatricia S. LanganSrikanta JanaRimas J. OrentasSheng Lin-GibsonWinfried KruegerBoro DropulićNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Barbara S. Paugh Lajos Baranyi Andre Roy Hua-Jun He Lindsay Harris Kenneth D. Cole Moria Artlip Caroline Raimund Patricia S. Langan Srikanta Jana Rimas J. Orentas Sheng Lin-Gibson Winfried Krueger Boro Dropulić Reference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells
description	Abstract Lentiviral vectors (LV) have emerged as a robust technology for therapeutic gene delivery into human cells as advanced medicinal products. As these products are increasingly commercialized, there are concomitant demands for their characterization to ensure safety, efficacy and consistency. Standards are essential for accurately measuring parameters for such product characterization. A critical parameter is the vector copy number (VCN) which measures the genetic dose of a transgene present in gene-modified cells. Here we describe a set of clonal Jurkat cell lines with defined copy numbers of a reference lentiviral vector integrated into their genomes. Genomic DNA was characterized for copy number, genomic integrity and integration coordinates and showed uniform performance across independent quantitative PCR assays. Stability studies during continuous long-term culture demonstrated sustained renewability of the reference standard source material. DNA from the Jurkat VCN standards would be useful for control of quantitative PCR assays for VCN determination in LV gene-modified cellular products and clinical samples.
format	article
author	Barbara S. Paugh Lajos Baranyi Andre Roy Hua-Jun He Lindsay Harris Kenneth D. Cole Moria Artlip Caroline Raimund Patricia S. Langan Srikanta Jana Rimas J. Orentas Sheng Lin-Gibson Winfried Krueger Boro Dropulić
author_facet	Barbara S. Paugh Lajos Baranyi Andre Roy Hua-Jun He Lindsay Harris Kenneth D. Cole Moria Artlip Caroline Raimund Patricia S. Langan Srikanta Jana Rimas J. Orentas Sheng Lin-Gibson Winfried Krueger Boro Dropulić
author_sort	Barbara S. Paugh
title	Reference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells
title_short	Reference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells
title_full	Reference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells
title_fullStr	Reference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells
title_full_unstemmed	Reference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells
title_sort	reference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/3dd0bd30801246de9358d8be2c031336
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Reference standards for accurate validation and optimization of assays that determine integrated lentiviral vector copy number in transduced cells

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