Abstract
A process of producing a receptor in HEK-293 cells used for the drug discovery program at Pfizer Inc. has been successfully developed with a novel BelloCell bioreactor to replace the conventional 2-D cell culturing devices including Cell Factories and roller bottles. A single BelloCell-500 has produced >1.4 × 109 HEK-293 cells, which are equivalent to those produced by 12 roller bottles, with substantially easier operation, single inoculation, less inoculum, less medium consumption and better space utilization. The receptor expression levels are better than those obtained by the traditional process. 3.7 pmoles of radioligandY mg−1 protein were attained in the bioreactor compared to 2.3 pmoles of radioligandY mg−1 protein in roller bottles. This may be attributed to the three dimensional attachment during cell growth. A 92% cell recovery from the bioreactor has been attained using Acutase or Trypsin treatment followed by four washes. It has been proven to be a viable and efficient device to produce adherent cells and express target components of interest for drug discovery applications.
Keywords: BelloCell-500, Cell Factory, G-protein coupled receptor, HEK293, Roller bottle
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References
- Chang M.M. and Ho L. 2003. Novel animal cell culture device for mass production of proteins and viruses. Genetic Engineering News, September 1.
- Hu Y.C., Lu J.T., Chung Y.C. High density cultivation of insect cells and production of recombinant baculovirus using a novel oscillating bioreactor. Cytotechnology. 2003;42:145–153. doi: 10.1023/B:CYTO.0000015841.98225.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Luo J., Yang S.-T. Effects of three-dimensional culturing in a fibrous matrix on cell cycleapoptosis and Mab production by hybridoma cells. Biotechnol. Prog. 2004;20:306–315. doi: 10.1021/bp034181v. [DOI] [PubMed] [Google Scholar]
- Mendonca R.Z., Pereira C.A. Cell metabolism and medium perfusion in Vero cell cultures on microcarriers in a bioreactor. Bioprocess Eng. 1998;18:213–218. doi: 10.1007/s004490050433. [DOI] [Google Scholar]
- Seeger T.F., Seymour P.A., Schmidt A.W., Zorn S.H., Schulz D.W., Lebel L.A., McLean S., Guanowsky V., Howard H.R., Lowe J.A., Heym J. Ziprasidone (CP-88,059): a new antipsychotic with combined dopamine and serotonin receptor antagonist activity. J. Pharmacol. Exp. Ther. 1995;275:101–113. [PubMed] [Google Scholar]
- Wang Y.C., Chang K.M., Lin F.L., Wu M.L. and Ho L. 2002. Novel disposable bioreactor for mammalian cell cultures and virus production. Poster in BioPHEX October 2002.