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Continuous production process of retroviral vector for adoptive T- cell therapy

Inwood, Sarah, Xu, Hui, Black, Mary A., Betenbaugh, Michael J., Feldman, Steven, Shiloach, Joseph
Biochemical engineering journal 2018 v.132 pp. 145-151
T-lymphocytes, antigens, bioreactors, bottles, dissolved oxygen, fibroblasts, mice, neoplasms, nutrients, pH, patients, receptors, retroviral vectors, therapeutics
Adoptive T-cell therapy is being considered as a promising method for cancer treatment. In this approach, patient’s T cells are isolated, modified, expanded, and administered back to the patient. Modifications may include adding specific T cell receptors (TCR) or chimeric antigen receptors (CAR) to the isolated cells by using retroviral vectors. PG13 cells, derivatives of NIH3T3 mouse fibroblasts, are being used to stably produce retroviral vectors that transduce the T cells. PG13 cells are anchorage-dependent cells that grow in roller bottles or cell factories and lately also in fixed bed bioreactors to produce the needed viral vector. To scale up viral vector production, PG13 cells were propagated on microcarriers in a stirred tank bioreactor utilizing an alternating tangential flow perfusion system. Microcarriers are 10 μm–0.5 mm beads that support the attachment of cells and are suspended in the bioreactor that provides controlled growth conditions. As a result, growth parameters, such as dissolved oxygen concentration, pH, and nutrients are monitored and continuously controlled. There were no detrimental effects on the specific viral vector titer or on the efficacy of the vector in transducing the T cells of several patients. Viral vector titer increased throughout the 11 days perfusion period, a total of 4.8 × 1011 transducing units (TU) were obtained with an average titer of 4.4 × 107 TU/mL and average specific productivity of 10.3 (TU) per cell, suggesting that this method can be an efficient way to produce large quantities of active vector suitable for clinical use.