Abstract
Medium optimization is an important strategy that can lead to several fold increase in the production of proteins in cell culture. However, the usual methods of medium optimization are complex and time consuming. Urokinase is a widely employed thrombolytic drug for the treatment of stroke. We describe here medium optimization for maximizing urokinase production by HT-1080 cells using supplementation of specific amino acids. The new specifically designed method resulted in 240 % increase in urokinase productivity.
References
Bedard C, Tom R, Kamen A (1993) Growth, nutrient consumption and end product accumulation in Sf-9 and BTI-EAA insect cell cultures: insights into growth limitation and metabolism. Biotechnol Prog 9(6):615–624
Chang JY, Knecht R, Braun DG (1981) Amino acid analysis at the picomole level. application to the C-terminal sequence analysis of polypeptides. Biochem J 199:547–555
Khaparde SS (2006) Urokinase production by animal cell culture in hollow fiber reactor, Ph.D. thesis, Indian Institute of Technology, New Delhi
Khaparde SS, Roychoudhury PK (2012) Effect of low culture temperature on urokinase production in hollow fiber reactor. Appl Biochem and Biotechnol 168(6):1655–1663. doi:10.1007/s12010-012-9886-2
Khaparde SS, Roychoudhury PK, Gomes J, Mukhopadhyay A (2008) External modulation of HT-1080 human fibrosarcoma cells improves urokinase production. Biotechnol Prog 24:1325–1332
Kuo M (1980) Urokinase production. US Patent 4,190,708
Lewis JL (1976) Production of urokinase. US Patent 3,930,945
Stoll TS, Mühlethaler K, von Stockar U, Marison IW (1996) Systematic improvement of a chemically defined protein free medium for hybridoma growth and monoclonal antibody production. J Biotechnol 45:111–123
Tait JF, Engelhardt S, Smith C, Fujikawa K (1995) Prourokinase-annexin V chimeras. J Biol Chem 37:21594–21599
Xing Z, Kenty B, Koyrakh I, Borys M, Pan S, Li Z (2011) Optimizing amino acid composition of CHO cell culture media for a fusion protein production. Process Biochem 46(7):1423–1429
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Khaparde, S.S., Roychoudhury, P.K. Amino acid supplementation enhances urokinase production by HT-1080 cells. J Ind Microbiol Biotechnol 41, 1035–1038 (2014). https://doi.org/10.1007/s10295-014-1434-3
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DOI: https://doi.org/10.1007/s10295-014-1434-3