Effects of Proline and Gelatin on hCTLA4Ig Production in Transgenic Rice Suspension Cells
2009
Song, M.N., Inha University, Incheon, Republic of Korea | Cheon, S.H., Inha University, Incheon, Republic of Korea | Kwon, J.Y., Inha University, Incheon, Republic of Korea | Choi, S.H., Inha University, Incheon, Republic of Korea | Kim, D.I., Inha University, Incheon, Republic of Korea
Rice cells were transformed to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter. hCTLA4Ig was produced and secreted into culture media inducibly when sugar was depleted. The obstacles of this system are the cell death and release of proteases by sugar starvation. These problems resulted in the losses of stability and productivity of hCTLA4Ig. Therefore, the effects of proline as an inhibitor of cell death were investigated. When 4 mM proline was added in sugar-free media, the cell death and release of proteases were reduced. As a consequence, the production of hCTLA4Ig was enhanced. In addition, the effects of protein stabilizers such as gelling agents were studied. It was found that the application of 0.01 g/L gelatin led to an increase in hCTLA4Ig production. This increase might be originated from the stabilization of hCTLA4Ig. In conclusion, the production of hCTLA4Ig could be enhanced by the additions of proline and gelatin in transgenic rice cell cultures.
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