full issue - Association of Biotechnology and Pharmacy
full issue - Association of Biotechnology and Pharmacy
full issue - Association of Biotechnology and Pharmacy
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Current Trends in <strong>Biotechnology</strong> <strong>and</strong> <strong>Pharmacy</strong><br />
Vol. 5 (3) 1298 -1308 July 2011, ISSN 0973-8916 (Print), 2230-7303 (Online)<br />
1306<br />
Activity <strong>of</strong> rhFVIII reached approximately<br />
7% after 120 h <strong>of</strong> cultivation when compared to<br />
that <strong>of</strong> the pooled plasma, while this amount was<br />
0% <strong>and</strong> 1.2% at 48 h <strong>and</strong> 96 h <strong>of</strong> cultivation,<br />
respectively. These results demonstrated that<br />
there is a direct relationship between protein<br />
production <strong>and</strong> cell density. Although there is<br />
no report on rhFVIII production in packed-bed<br />
bioreactors in the literature, but they have been<br />
employed for production <strong>of</strong> some recombinant<br />
pharmaceuticals (4). Trombopoietin was<br />
produced by the rCHO cells attached to polyester<br />
microcarriers in a packed-bed bioreactor. The<br />
maximum cell density <strong>and</strong> productivity <strong>of</strong><br />
2×10 7 cells/ml <strong>and</strong> 1.3-1.8 mg/l.d were obtained<br />
after 15 days, respectively (Cong et al. 2001).<br />
Because <strong>of</strong> the low stability <strong>of</strong> rhFVIII, the<br />
culture time period should be decreased. The<br />
results show that rhFVIII is biologically active<br />
during 120 h <strong>of</strong> cultivation when using enriched<br />
medium, in the packed bed bioreactor.<br />
Conclusion<br />
To our knowledge, no reports have been<br />
found in the literature on the physico-chemical<br />
culture conditions <strong>of</strong> recombinant CHO cells for<br />
improvement <strong>of</strong> rhFVIII production. The results<br />
<strong>of</strong> this study have demonstrated that medium<br />
supplemented with amino acids, precursors <strong>and</strong><br />
glucose has a significant positive effect on the<br />
rCHO cell density <strong>and</strong> rhFVIII production,<br />
mainly because <strong>of</strong> the reduction <strong>of</strong> by-products<br />
formation. These data have provided useful<br />
information for the process development. Since<br />
rhFVIII is very sensitive to proteolytic<br />
degradation, culture time is an important factor<br />
in achieving an appropriate rhFVIII activity.<br />
Therefore, continuous packed-bed or perfusion<br />
cultures are suggested as suitable systems for<br />
production <strong>of</strong> rhFVIII in large-scale cultivation.<br />
Acknowledgments<br />
This work was financially supported in<br />
part by the International Center for Genetic<br />
Engineering <strong>and</strong> <strong>Biotechnology</strong> (ICGEB, Italy).<br />
The authors thank Dr. Parvin Shariati for her<br />
great assistance.<br />
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