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.
Similar content being viewed by others
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.
Y.C. Hu J.T. Lu Y.C. Chung (2003) ArticleTitleHigh density cultivation of insect cells and production of recombinant baculovirus using a novel oscillating bioreactor Cytotechnology 42 145–153 Occurrence Handle10.1023/B:CYTO.0000015841.98225.27
J. Luo S.-T. Yang (2004) ArticleTitleEffects of three-dimensional culturing in a fibrous matrix on cell cycleapoptosis and Mab production by hybridoma cells Biotechnol. Prog. 20 306–315 Occurrence Handle10.1021/bp034181v Occurrence Handle14763857
R.Z. Mendonca C.A. Pereira (1998) ArticleTitleCell metabolism and medium perfusion in Vero cell cultures on microcarriers in a bioreactor Bioprocess Eng. 18 213–218 Occurrence Handle10.1007/s004490050433
T.F. Seeger P.A. Seymour A.W. Schmidt S.H. Zorn D.W. Schulz L.A. Lebel S. McLean V. Guanowsky H.R. Howard J.A. Lowe J. Heym (1995) ArticleTitleZiprasidone (CP-88,059): a new antipsychotic with combined dopamine and serotonin receptor antagonist activity J. Pharmacol. Exp. Ther. 275 101–113 Occurrence Handle7562537
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ho, L., Greene, C.L., Schmidt, A.W. et al. Cultivation of HEK 293 cell line and production of a member of the superfamily of G-protein coupled receptors for drug discovery applications using a highly efficient novel bioreactor. Cytotechnology 45, 117–123 (2004). https://doi.org/10.1007/s10616-004-6402-8
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10616-004-6402-8