Frankfurt am Main, Germany
Frankfurt am Main, Germany

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Loew R.,EUFETS AG | Meyer Y.,EUFETS AG | Kuehlcke K.,EUFETS AG | Gama-Norton L.,Helmholtz Center for Infection Research | And 9 more authors.
Gene Therapy | Year: 2010

The clinical application of self-inactivating (SIN) retroviral vectors has been hampered by the lack of reliable and efficient vector production technologies. To enable production of SIN γ-retroviral vectors from stable producer clones, a new PG13-based packaging cell, known as PG368, was developed. Viral vector expression constructs can be reliably inserted at a predefined genomic locus of PG368 packaging cells by an Flp-recombinase-mediated targeted cassette exchange (RMCE) reaction. A new, carefully designed vector-targeting construct, pEMTAR-1, eliminated the co-packaging of the selectable marker gene used for the identification of successful recombination at the predefined genomic locus and thus, improved the safety of the production system. Selected clones produced vector supernatants at consistent titers. The targeted insertion of therapeutically relevant SIN vectors for chronic granulomatous disease and X-linked severe combined immunodeficiency into PG368 cells results in stable titers within the range necessary for clinical application. The production of retroviral SIN vectors from stable clinical-grade producer cells is feasible and will contribute to the safe production and application of SIN γ-retroviral vectors for clinical trials. © 2010 Macmillan Publishers Limited. All rights reserved.

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