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Effect of Osmotic Pressure on hCTLA-lg Production in Transgenic Rice Cell Suspension Cultures  

Choi Sung-Hun (Department of Biological Engineering, Inha University)
Lee Song-Jae (Boryung Central Research institute, Boryung Pharmaceutical Co. Ltd.)
Hong Seok-Mi (Department of Biological Engineering, Inha University)
Cho Ji-Suk (Department of Biological Engineering, Inha University)
Kim Dong-Il (Department of Biological Engineering, Inha University)
Publication Information
KSBB Journal / v.20, no.4, 2005 , pp. 278-284 More about this Journal
Abstract
An immunosuppressive agent, human cytotoxic T lymphocyte antigen 4 (hCTLA4), is used for the prevention of graft rejection and treatment of autoimmune diseases. hCTLA4-Ig, a CTLA4-immunoglobulin fusion protein, was produced and secreted from transgenic rice cell suspension cultures using rice a-amylase (RAmy3D) expression system. In this system, hCTLA4-Ig expression was regulated metabolically by sugar starvation. For the purpose of improving hCTLA4-Ig production, the effects of osmotic pressure was investigated in suspension cultures of transgenic rice cells. The highest production level was achieved at 40 mM sorbitol $(140\;mOsm{\cdot}kg^{-1}\;H_2O)$. Using the medium with 8 mM glucose, the level of hCTLA4-Ig in the medium reached 45.3 mg/L. By adjusting the osmotic pressure of induction medium, it was found that the hCTLA4-Ig production could be increased up to 2.1-fold compared with that in batch culture.
Keywords
Osmotic pressure; sorbitol; RAmy3D promoter; Oryza sativa L.; hCTLA4-lg;
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