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Abstract
The compelling need to provide adoptive cell therapy (ACT) to an increasing number of oncology patients within a meaningful therapeutic window makes the development of an efficient, fast, versatile, and safe genetic tool for creating recombinant T cells indispensable. In this study, we used nonintegrating minimally sized DNA vectors with an enhanced capability of generating genetically modified cells, and we demonstrate that they can be efficiently used to engineer human T lymphocytes. This vector platform contains no viral components and is capable of replicating extrachromosomally in the nucleus of dividing cells, providing persistent transgene expression in human T cells without affecting their behavior and molecular integrity. We use this technology to provide a manufacturing protocol to quickly generate chimeric antigen receptor (CAR)–T cells at clinical scale in a closed system and demonstrate their enhanced anti-tumor activity in vitro and in vivo in comparis ....

Amersham Bioscience , D Trono Addgene , Texmacs Miltenyi Biotec , Miltenyi Biotec , Lumikine Xpress , Jackson Immunoresearch , Maxcyte Gtx , Panreac Applichem , Deutsches Krebsforschungszentrum , Elevator Pitch Programme Of The National Center , Bd Sciences , Drug Administration , Agilent Technologies , Proof Of Concept Program , Deutsches Krebsforschungszentrum German Cancer Research Center , Nature Technology Corporation , Jackson Laboratory , Heidelberg University Hospital Blood Bank , Stellar Competent Cells , Life Technologies , Leinco Technologies , Acro Biosystems , Alexa Fluor , Tissue Extraction , Thermo Fisher Scientific , Loading Dye ,