Clinical grade vector production: analysis of yield, stability, and storage of gmp-produced retroviral vectors for gene therapy.
Retroviral vector gene transfer of a therapeutic gene to correct or modify a disease process is a promising strategy for many inherited and acquired diseases. A major obstacle in this process is the large-scale production of the gene transfer vector under good manufacturing practice (GMP) conditions. We have used the CellCube bioreactor system to produce five batches of GMP-grade vector. The production batches were of 10-20 L each, and the titers were around 2 x 10(6) IU/mL. We find that this particular vector is relatively stable with a half-life of about 8 h at 37 degrees C, 40 h at 20 degrees C, and 14 days at 4 degrees C. The half-life during storage at -80 degrees C is around 18 months. The supernatant may be frozen and thawed up to five times without any significant loss of titer. We have also made a comparison between the CellCube bioreactor and the automated roller bottle system RollerCell 40 (RC 40). The yields from the two systems were comparable.
Wikström
Kristina
K
Vecura, The Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, SE-141 86 Stockholm, Sweden. kristina.wikstrom@crc.ki.se
Blomberg
Pontus
P
Islam
Khalid B
KB
D001483:Base Sequence; D017931:DNA Primers; D015316:Genetic Therapy; D005822:Genetic Vectors; D012190:Retroviridae / Q000235:genetics*
D017931:DNA Primers