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.
Vecura, The Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, SE-141 86 Stockholm, Sweden. kristina.wikstrom@crc.ki.se
Kristina
K
Wikström
Pontus
P
Blomberg
Khalid B
KB
Islam
D001483:Base Sequence; D017931:DNA Primers; D015316:Genetic Therapy; D005822:Genetic Vectors; D012190:Retroviridae / Q000235:genetics*
D017931:DNA Primers