A Statistical Experimental Design for the Enhanced Production of hGMCSF

초록

Transgenic plant cell suspension cultures have been considered as an effective alternative system to produce recombinant proteins for industrial and pharmaceutical uses. Rice (Oryza sativa L.) cells were genetically modified to express human granulocyte-macrophage colony-stimulating factor (hGM-CSF). hGM-CSF is one of 4 specific glycoproteins that stimulate a population of committed granulocyte macrophage progenitor cells to generate granulocytes and macrophages. hGM-CSF was produced with a rice α-amylase (RAmy3D) promoter system and secreted into culture media when carbon sources were depleted at stationary phase. Therefore, the culture period can be divided into cell growth phase and production phase. In this study, a statistical experimental design were used to optimize media compositions and batch culture conditions. The cell growth and hGM-CSF production could be improved. Plackett-Burman and Box-Behnken design with Minitab software were applied for screening of main factors and determining of optimal levels. Main factors were studied sucrose, nitrogen, phosphate 2,4-D, kinetin and temperature. The statistical analysis of factorial effects resulted in three important parameters, which were sucrose, 2,4-D and kinetin.