2.50
Hdl Handle:
http://hdl.handle.net/10033/141114
Title:
Integrated strategy for the production of therapeutic retroviral vectors.
Authors:
Carrondo, Manuel; Panet, Amos; Wirth, Dagmar; Coroadinha, Ana Sofia; Cruz, Pedro; Falk, Haya; Schucht, Roland; Dupont, Francis; Geny-Fiamma, Cécile; Merten, Otto-Wilhelm; Hauser, Hansjörg
Abstract:
The broad application of retroviral vectors for gene delivery is still hampered by the difficulty to reproducibly establish high vector producer cell lines generating sufficient amounts of highly concentrated virus vector preparations of high quality. To enhance the process for producing clinically relevant retroviral vector preparations for therapeutic applications, we have integrated novel and state-of-the-art technologies in a process that allows rapid access to high-efficiency vector-producing cells and consistent production, purification, and storage of retroviral vectors. The process has been designed for various types of retroviral vectors for clinical application and to support a high-throughput process. New modular helper cell lines that permit rapid insertion of DNA encoding the therapeutic vector of interest at predetermined, optimal chromosomal loci were developed to facilitate stable and high vector production levels. Packaging cell lines, cultivation methods, and improved medium composition were coupled with vector purification and storage process strategies that yield maximal vector infectivity and stability. To facilitate GMP-grade vector production, standard of operation protocols were established. These processes were validated by production of retroviral vector lots that drive the expression of type VII collagen (Col7) for the treatment of a skin genetic disease, dystrophic epidermolysis bullosa. The potential efficacy of the Col7-expressing vectors was finally proven with newly developed systems, in particular in target primary keratinocyte cultures and three-dimensional skin tissues in organ culture.
Affiliation:
Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, P-2781-901 Oeiras, Portugal.
Citation:
Integrated strategy for the production of therapeutic retroviral vectors. 2011, 22 (3):370-9 Hum. Gene Ther.
Journal:
Human gene therapy
Issue Date:
Mar-2011
URI:
http://hdl.handle.net/10033/141114
DOI:
10.1089/hum.2009.165
PubMed ID:
21043806
Type:
Article
Language:
en
ISSN:
1557-7422
Appears in Collections:
Publications of Dept. Gene Regulation and Differentiation (RDIF)

Full metadata record

DC FieldValue Language
dc.contributor.authorCarrondo, Manuelen
dc.contributor.authorPanet, Amosen
dc.contributor.authorWirth, Dagmaren
dc.contributor.authorCoroadinha, Ana Sofiaen
dc.contributor.authorCruz, Pedroen
dc.contributor.authorFalk, Hayaen
dc.contributor.authorSchucht, Rolanden
dc.contributor.authorDupont, Francisen
dc.contributor.authorGeny-Fiamma, Cécileen
dc.contributor.authorMerten, Otto-Wilhelmen
dc.contributor.authorHauser, Hansjörgen
dc.date.accessioned2011-08-29T12:31:56Z-
dc.date.available2011-08-29T12:31:56Z-
dc.date.issued2011-03-
dc.identifier.citationIntegrated strategy for the production of therapeutic retroviral vectors. 2011, 22 (3):370-9 Hum. Gene Ther.en
dc.identifier.issn1557-7422-
dc.identifier.pmid21043806-
dc.identifier.doi10.1089/hum.2009.165-
dc.identifier.urihttp://hdl.handle.net/10033/141114-
dc.description.abstractThe broad application of retroviral vectors for gene delivery is still hampered by the difficulty to reproducibly establish high vector producer cell lines generating sufficient amounts of highly concentrated virus vector preparations of high quality. To enhance the process for producing clinically relevant retroviral vector preparations for therapeutic applications, we have integrated novel and state-of-the-art technologies in a process that allows rapid access to high-efficiency vector-producing cells and consistent production, purification, and storage of retroviral vectors. The process has been designed for various types of retroviral vectors for clinical application and to support a high-throughput process. New modular helper cell lines that permit rapid insertion of DNA encoding the therapeutic vector of interest at predetermined, optimal chromosomal loci were developed to facilitate stable and high vector production levels. Packaging cell lines, cultivation methods, and improved medium composition were coupled with vector purification and storage process strategies that yield maximal vector infectivity and stability. To facilitate GMP-grade vector production, standard of operation protocols were established. These processes were validated by production of retroviral vector lots that drive the expression of type VII collagen (Col7) for the treatment of a skin genetic disease, dystrophic epidermolysis bullosa. The potential efficacy of the Col7-expressing vectors was finally proven with newly developed systems, in particular in target primary keratinocyte cultures and three-dimensional skin tissues in organ culture.en
dc.language.isoenen
dc.subject.meshAnimalsen
dc.subject.meshCell Culture Techniquesen
dc.subject.meshCell Lineen
dc.subject.meshCollagen Type VIIen
dc.subject.meshDNA Nucleotidyltransferasesen
dc.subject.meshEpidermolysis Bullosa Dystrophicaen
dc.subject.meshGene Therapyen
dc.subject.meshGenetic Vectorsen
dc.subject.meshHEK293 Cellsen
dc.subject.meshHumansen
dc.subject.meshIndustrial Microbiologyen
dc.subject.meshMiceen
dc.subject.meshMice, Inbred BALB Cen
dc.subject.meshRecombination, Geneticen
dc.subject.meshReproducibility of Resultsen
dc.subject.meshRetroviridaeen
dc.titleIntegrated strategy for the production of therapeutic retroviral vectors.en
dc.typeArticleen
dc.contributor.departmentInstituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, P-2781-901 Oeiras, Portugal.en
dc.identifier.journalHuman gene therapyen

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