Improvement of rCHO Cell Density and Heterologous Protein Production: Effect of Culture Conditions

Order of Publishing in Issue: 
Volume :5
Issue :3
July, 2011
Page No: 
1298 -1308
M. Shakibaie [1,2], F. Tabandeh [1]*, A. Zomorodipour [3], S. Ebrahimi [2], F. Ataie [3] and H. Habib Ghomi [1]
[1] Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, P.O. Box: 14965-161, Tehran, Iran
[2] Biotechnology Group, Chemical Engineering Faculty, Sahand University of Technology, Tabriz, Iran
[3] Department of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology, P.O. Box: 14965-161, Tehran, Iran

Abstract : Physico-chemical parameters have a great impact on mammalian cell growth. In this study, the effects of a number of amino acids,precursors, serum, glucose, osmolality and temperature on the recombinant Chinese hamster ovary (rCHO) cells producing the recombinanthuman coagulation factor VIII (rhFVIII) were investigated, so as to improve the rCHO cell density in the packed-bed bioreactor. The resultsshowed that while the concentration of ammonium, as the main by-product, increased by more than 6mM, the rCHO cell densitydecreased significantly. On the other hand, adding certain amino acids and precursors mitigated the stress caused by ammonium andconsequently increased cell density. Proline and glycine increased cell density up to approximately 28% in comparison with thenegative control. Nucleic acids, as the main precursors, had a significant impact on rCHO cell growth, which increased by up to 20%.Higher amounts of glucose (3.5 g/l) supplied the carbon and energy requirements of the cells, inhibited by-products formation and sequentiallyincreased heterologous protein production. The precursors allowed a 14% improvement in rhFVIII activity at high glucose concentrations,which was 1.5 fold greater than the negative control. Both hyperosmotic pressure and temperature downshifts had remarkable adverseeffects on the rCHO cell density,; being reduced to 2.7×106 cells/ml. The acquired suitable culture conditions were applied to the packed-bedbioreactor, and consequently an rCHO cell density of 18×108 cells/l was achieved during 120 h of cultivation. The results of this studyindicated that the rCHO cells essentially need supplementary materials to deal with inappropriate culture conditions; especially in aserum-free medium.

Factor VIII, Packed-bed bioreactor, Recombinant CHO cells, Serum-free medium.
Paper-6.pdf920.14 KB

To download full text articles and issues in PDF at no cost.All you have to do is fill out a brief registration form. It's easy and it's free. At any time, you may request removal of your contact information from our database.
Click here to Register | Click here if you have already registered

2018 ABAP Awards

Invitation for Nominations to 2019 ABAP Awards
The Association of Biotechnology and Pharmacy invites nominations for the following awards from the eligible talented people with proven track record and achievements on or before August 31, 2019 in the prescribed format.



User login