Clinical scale production of an improved retroviral vector expressing the human multidrug resistance 1 gene (MDR1).
Retroviral vectors are currently the most important and best characterized tools for ex vivo genetic modification of hematopoietic progenitor/stem cells. As a prerequisite for clinical applications, large volumes of high-titer vector supernatants have to be generated in compliance with 'GMP' guidelines. This goal can be reached using a carefully selected producer cell clone and a conventional large-scale cell culture system. The retroviral vector SF1m provides efficient expression of the human multidrug resistance 1 (MDR1) gene in hematopoietic progenitor/stem cells in vitro and in NOD/SCID mouse repopulating human cells in vivo. Currently, a clinical phase I/II study is in preparation to test whether intensified consolidation chemotherapy is enabled by autologous transplantation of peripheral blood progenitor/stem cells that have been genetically modified with SF1m. Using multi-tray cell factories >19 l of serum-free vector containing supernatant were generated from cells of a previously established SF1m-producer clone, based on the PG13 packaging cell line. Testing of the final samples revealed sufficient quality (>1.5 x 10(6) infectious particles/ml) for clinical scale transduction of CD34+ cells. Results from the production runs and the applied biosafety concept are described.
Eckert
H G
HG
EUFETS GmbH, Idar-Oberstein, Germany.
Kühlcke
K
K
Schilz
A J
AJ
Lindemann
C
C
Basara
N
N
Fauser
A A
AA
Baum
C
C
D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage; D001709:Biotechnology; D002460:Cell Line; D019450:Genes, MDR; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009369:Neoplasms / Q000188:drug therapy / Q000628:therapy; D012190:Retroviridae / Q000235:genetics*; D012449:Safety; D014182:Transplantation, Autologous
D000970:Antineoplastic Agents