Large scale production of recombinant mouse and rat growth hormone by fed-batch GS-NSO cell cultures.

Cytotechnology
W ZhouJ Aunins

Abstract

Investigations of biological effects of prolonged elevation of growth hormone in animals such as mice and rats require large amounts of mouse and rat growth hormone (GH) materials. As an alternative to scarce and expensive pituitary derived materials, both mouse and rat GH were expressed in NSO murine myeloma cells transfected with a vector containing the glutamine synthetase (GS) gene and two copies of mouse or rat GH cDNA. For optimal expression, the mouse GH vector also contained sequences for targeting integration by homologous recombination. Fed-batch culture processes for such clones were developed using a serum-free, glutamine-free medium and scaled up to 250 L production scale reactors. Concentrated solutions of proteins, amino acids and glucose were fed periodically to extend cell growth and culture lifetime, which led to an increase in the maximum viable cell concentration to 3.5×10(9) cells/L and an up to 10 fold increase in final mouse and rat rGH titers in comparison with batch cultures. For successful scale up, similar culture environmental conditions were maintained at different scales, and specific issues in large scale reactors such as balancing oxygen supply and carbon dioxide removal, were addressed. Very sim...Continue Reading

Citations

Jul 19, 2008·Biotechnology and Bioengineering·W ZhouJ Aunins
Sep 6, 2001·Biotechnology and Bioengineering·J J OsmanJ Varley
Jul 26, 2000·Journal of Biotechnology·H J CruzM J Carrondo
Nov 13, 2008·Cytotechnology·J Varley, J Birch
Apr 15, 2005·Biotechnology and Bioengineering·Gargi SethWei-Shou Hu
Mar 21, 2007·Toxicological Sciences : an Official Journal of the Society of Toxicology·Georgia M FarrisSrinivasa Prahalada

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