Cargando…

Scalable continuous-flow electroporation platform enabling T cell transfection for cellular therapy manufacturing

Viral vectors represent a bottleneck in the manufacturing of cellular therapies. Electroporation has emerged as an approach for non-viral transfection of primary cells, but standard cuvette-based approaches suffer from low throughput, difficult optimization, and incompatibility with large-scale cell...

Descripción completa

Detalles Bibliográficos
Autores principales: VanderBurgh, Jacob A., Corso, Thomas N., Levy, Stephen L., Craighead, Harold G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10133335/
https://www.ncbi.nlm.nih.gov/pubmed/37185305
http://dx.doi.org/10.1038/s41598-023-33941-2
_version_ 1785031548199763968
author VanderBurgh, Jacob A.
Corso, Thomas N.
Levy, Stephen L.
Craighead, Harold G.
author_facet VanderBurgh, Jacob A.
Corso, Thomas N.
Levy, Stephen L.
Craighead, Harold G.
author_sort VanderBurgh, Jacob A.
collection PubMed
description Viral vectors represent a bottleneck in the manufacturing of cellular therapies. Electroporation has emerged as an approach for non-viral transfection of primary cells, but standard cuvette-based approaches suffer from low throughput, difficult optimization, and incompatibility with large-scale cell manufacturing. Here, we present a novel electroporation platform capable of rapid and reproducible electroporation that can efficiently transfect small volumes of cells for research and process optimization and scale to volumes required for applications in cellular therapy. We demonstrate delivery of plasmid DNA and mRNA to primary human T cells with high efficiency and viability, such as > 95% transfection efficiency for mRNA delivery with < 2% loss of cell viability compared to control cells. We present methods for scaling delivery that achieve an experimental throughput of 256 million cells/min. Finally, we demonstrate a therapeutically relevant modification of primary T cells using CRISPR/Cas9 to knockdown T cell receptor (TCR) expression. This study displays the capabilities of our system to address unmet needs for efficient, non-viral engineering of T cells for cell manufacturing.
format Online
Article
Text
id pubmed-10133335
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-101333352023-04-28 Scalable continuous-flow electroporation platform enabling T cell transfection for cellular therapy manufacturing VanderBurgh, Jacob A. Corso, Thomas N. Levy, Stephen L. Craighead, Harold G. Sci Rep Article Viral vectors represent a bottleneck in the manufacturing of cellular therapies. Electroporation has emerged as an approach for non-viral transfection of primary cells, but standard cuvette-based approaches suffer from low throughput, difficult optimization, and incompatibility with large-scale cell manufacturing. Here, we present a novel electroporation platform capable of rapid and reproducible electroporation that can efficiently transfect small volumes of cells for research and process optimization and scale to volumes required for applications in cellular therapy. We demonstrate delivery of plasmid DNA and mRNA to primary human T cells with high efficiency and viability, such as > 95% transfection efficiency for mRNA delivery with < 2% loss of cell viability compared to control cells. We present methods for scaling delivery that achieve an experimental throughput of 256 million cells/min. Finally, we demonstrate a therapeutically relevant modification of primary T cells using CRISPR/Cas9 to knockdown T cell receptor (TCR) expression. This study displays the capabilities of our system to address unmet needs for efficient, non-viral engineering of T cells for cell manufacturing. Nature Publishing Group UK 2023-04-26 /pmc/articles/PMC10133335/ /pubmed/37185305 http://dx.doi.org/10.1038/s41598-023-33941-2 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
VanderBurgh, Jacob A.
Corso, Thomas N.
Levy, Stephen L.
Craighead, Harold G.
Scalable continuous-flow electroporation platform enabling T cell transfection for cellular therapy manufacturing
title Scalable continuous-flow electroporation platform enabling T cell transfection for cellular therapy manufacturing
title_full Scalable continuous-flow electroporation platform enabling T cell transfection for cellular therapy manufacturing
title_fullStr Scalable continuous-flow electroporation platform enabling T cell transfection for cellular therapy manufacturing
title_full_unstemmed Scalable continuous-flow electroporation platform enabling T cell transfection for cellular therapy manufacturing
title_short Scalable continuous-flow electroporation platform enabling T cell transfection for cellular therapy manufacturing
title_sort scalable continuous-flow electroporation platform enabling t cell transfection for cellular therapy manufacturing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10133335/
https://www.ncbi.nlm.nih.gov/pubmed/37185305
http://dx.doi.org/10.1038/s41598-023-33941-2
work_keys_str_mv AT vanderburghjacoba scalablecontinuousflowelectroporationplatformenablingtcelltransfectionforcellulartherapymanufacturing
AT corsothomasn scalablecontinuousflowelectroporationplatformenablingtcelltransfectionforcellulartherapymanufacturing
AT levystephenl scalablecontinuousflowelectroporationplatformenablingtcelltransfectionforcellulartherapymanufacturing
AT craigheadharoldg scalablecontinuousflowelectroporationplatformenablingtcelltransfectionforcellulartherapymanufacturing