TY - JOUR
T1 - Cost-effective manufacture of an allogeneic GM-CSF-secreting breast tumor vaccine in an academic cGMP facility
AU - Davis-Sproul, Janice M.
AU - Harris, M. P.
AU - Davidson, N. E.
AU - Kobrin, B. J.
AU - Jaffee, E. M.
AU - Emens, L. A.
N1 - Funding Information:
We thank Timothy Hansen and Lynn Santarsiero for technical assistance, and Renee Porter for quality assurance. We also thank the manufacturing and regulatory staff of the Cell Processing and Gene Therapy Facility at the Johns Hopkins University. This work is supported in part by the Department of Defense grant DAMD17-01-1-0281 (to LA Emens), the Maryland Cigarette Restitution Fund M020216 (to LA Emens), the SPORE in Breast Cancer Grant 1P50CA88843 (to LA Emens, EM Jaffee and NE Davidson), the Breast Cancer Research Foundation (to EM Jaffee and NE Davidson), and the AVON Foundation Baltimore/Seattle Immunotherapy Collaborative Grant (to EM Jaffee). EM Jaffee is the first recipient of the Dana and Albert ‘Cubby’ Broccoli Professorship in Oncology.
PY - 2005
Y1 - 2005
N2 - Background: GM-CSF-secreting, allogeneic cell-based cancer vaccines have shown promise for the treatment of a variety of solid tumors. We have now applied this approach to breast cancer. The aim of these studies was to optimize expansion parameters, qualify the manufacturing process, and establish expected outcomes for cGMP-compliant manufacturing of two GM-CSF-secreting breast tumor cell lines. Methods: The variables affecting the efficiency of expanding and formulating two allogeneic GM-CSF-secreting cell lines, 2T47D-V and 3SKBR3-7, were systematically evaluated. Production criteria investigated included alternative cell culture vessels (flasks vs. cell factories), centrifugation time and speed variables for large volume cell concentration, cell seeding density, the minimal concentration of FBS required for maximal cell expansion, and the dose and timing of irradiation in relation to cryopreservation. Results: These studies demonstrate that, in comparison with standard 150-cm2 tissue culture flasks, Nunc 10-Stack Cell Factories are a more efficient and practical cell culture vessel for vaccine cell line manufacture. Centrifugation optimization studies using the COBE® 2991™ Cell Processor established that a speed of 2000 r.p.m. (450 g) for 2 min reliably concentrated the cells while maintaining acceptable viability and bioactivity. Radiation studies established that lethal irradiation prior to cryopreservation does not compromise the quality of the product, as measured by post-thaw cell viability and GM-CSF cell line-specific secretion levels. Finally, studies aimed at optimizing the production of one vaccine cell line, 3SKBR3-7, demonstrated that seeding the cells at a higher density and maintaining them in half the initial concentration of FBS maximized the yield of bioactive cells, resulting in significant cost savings. Discussion: A manufacturing process that simultaneously maximizes cell yield, minimizes cell manipulation and maintains vaccine cell potency is critical for producing cell-based cancer vaccines in an academic setting. These studies define a feasible, reproducible and cost-effective methodology for production of a GM-CSF-secreting breast cancer vaccine that is cGMP compliant.
AB - Background: GM-CSF-secreting, allogeneic cell-based cancer vaccines have shown promise for the treatment of a variety of solid tumors. We have now applied this approach to breast cancer. The aim of these studies was to optimize expansion parameters, qualify the manufacturing process, and establish expected outcomes for cGMP-compliant manufacturing of two GM-CSF-secreting breast tumor cell lines. Methods: The variables affecting the efficiency of expanding and formulating two allogeneic GM-CSF-secreting cell lines, 2T47D-V and 3SKBR3-7, were systematically evaluated. Production criteria investigated included alternative cell culture vessels (flasks vs. cell factories), centrifugation time and speed variables for large volume cell concentration, cell seeding density, the minimal concentration of FBS required for maximal cell expansion, and the dose and timing of irradiation in relation to cryopreservation. Results: These studies demonstrate that, in comparison with standard 150-cm2 tissue culture flasks, Nunc 10-Stack Cell Factories are a more efficient and practical cell culture vessel for vaccine cell line manufacture. Centrifugation optimization studies using the COBE® 2991™ Cell Processor established that a speed of 2000 r.p.m. (450 g) for 2 min reliably concentrated the cells while maintaining acceptable viability and bioactivity. Radiation studies established that lethal irradiation prior to cryopreservation does not compromise the quality of the product, as measured by post-thaw cell viability and GM-CSF cell line-specific secretion levels. Finally, studies aimed at optimizing the production of one vaccine cell line, 3SKBR3-7, demonstrated that seeding the cells at a higher density and maintaining them in half the initial concentration of FBS maximized the yield of bioactive cells, resulting in significant cost savings. Discussion: A manufacturing process that simultaneously maximizes cell yield, minimizes cell manipulation and maintains vaccine cell potency is critical for producing cell-based cancer vaccines in an academic setting. These studies define a feasible, reproducible and cost-effective methodology for production of a GM-CSF-secreting breast cancer vaccine that is cGMP compliant.
KW - Allogeneic cell lines
KW - Cancer vaccine
KW - Cell factory
KW - GM-CSF
KW - cGMP
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U2 - 10.1080/14653240510018082
DO - 10.1080/14653240510018082
M3 - Article
C2 - 16040383
AN - SCOPUS:17444410345
SN - 1465-3249
VL - 7
SP - 46
EP - 56
JO - Cytotherapy
JF - Cytotherapy
IS - 1
ER -