Enhanced Production of hCTLA4Ig in Transgenic Rice Cell Suspension Cultures by Using Pluronic Surfactants

초록

Genetically-modified cells of Oryza sativa L. cv. Dongjin were cultivated to produce hCTLA4Ig using RAmy3D promoter. Stability of hCTLA4Ig was affected by various kinds of proteases that were related to stress-induced cell suicide. This proteolytic degradation obstructed the high-level production of hCTLA4Ig. The objective of this study was to protect the cells against stress and to decrease the loss of productivity. Pluronic surfactants were known to protect mammalian and insects cells from the damage caused by hydrodynamic shear stresses. To investigate the effects of various Pluronic surfactants on transgenic rice cell cultures, Pluronic F-68, Pluronic F-127, Plurafac A-38 and Pluronic 10R5 were tested. Production of hCTLA4Ig at various concentrations (0.001, 0.01, 0.1, 1, 3, 5 and 7 g/L) was compared with that in control cultures. It was found that the addition of 1 g/L Pluronic 10R5 led to higher hCTLA4Ig productivity than control cultures. The maximum hCTLA4Ig concentration was 32 mg/L with 1 g/L Pluronic 10R5 compared to 22 mg/L .