Long-Term CB.Hep-1 Monoclonal Antibody Production in a Single-Use Bioreactor on a Rocker Platform with Serum- and Protein-Free Media

by Alberto Galván, Rodolfo Valdés, Marcos González, Hasel Aragón, Daily Hernández, Sigifredo Padilla, Leonardo Gómez, David Gailán, Yanet Villegas, Andrés Tamayo, Adelma Pérez, Bárbara Pérez, Maylín Lao, Yodelis Calvo, Aneet Fernández, Lianys Lee, Jania Suarez, Judey García, and Eduardo Ojito
Volume 14, Issue 4 (Winter 2015/2016)

Rocker bag bioreactors have been used successfully in cultivating cells because they provide good nutrient distribution and cell suspension while eliminating the need to validate cleaning and sterilization. Therefore, this study examined the long-term performance of a 50 L single-use bag bioreactor on a rocking platform in CB.Hep-1 monoclonal antibody (mAb) production. For such a purpose, the bioreactor was operated in a continuous mode with a mixture of serum-free media (SFM) for 62 days, and with protein-free medium (PFM) for another 62 days. Stationary phase culture results with SFM were: cell concentration of 1.57 ± 0.2 × 106 cells mL-1, specific growth rate of (μ)=0.0202, cell viability of 91.1 ± 6.4%, mAb concentration of 44.6 ± 11.1 μg mL-1, cell-specific secretion of 28.8 ± 8.1 pg cell-1 , and mAb yield of 42.4 ± 4.0 mg L-1. After SFM was replaced by PFM, there were statistically different results (p<0.05) in a cell concentration of 2.56 ± 0.2 × 106 cells mL-1, cell viability of 97.0 ± 1.1%, mAb concentration of 90.1 ± 28.2 μg mL-1, and mAb yield of 76.6 ± 10.6 mg L-1. However, the cell-specific secretion of 38.0 ± 6.0 pg cell-1 was statistically similar, but only with the first batch run with SFM. Average purification recovery from the SFM and PFM supernatants was 74.6 ± 7.9% and 70.9 ± 11.2%, respectively. The recovery and biochemical properties of the CB.Hep-1 mAb cultured with either media composition were similar to those found with CB.Hep-1 mAb produced by ascites. Regardless of whether SFM or PFM is used, it can be concluded that the 50 L single-use bag bioreactor on a rocker platform was suitable for long-term CB.Hep-1 production and can result in a 36 g yield in 125 days...

Galván A, Valdés R, González M, Aragón H, Hernández D, Padilla S et al. Long-term CB.Hep-1 monoclonal antibody production in a single-use bioreactor on a rocker platform with serum- and protein-free media. BioProcess J, 2016; 14(4): 4–13. http://dx.doi.org/10.12665/J144.Valdes.

Posted online January 12, 2016.