High-Efficiency Expression and Production System of Heterologous Protein in Transgenic Rice Cell Cultur (III)

Protein production by plant cell culture has a number of advantages for biotechnology application. Using plant cell to produce heterologous proteins has become important and valuable in both medical and industry. The research and development of scale-up production technology of high-value proteins, vaccine or antibody is the most important issue in the whole biotechnology fields. In this study, we will develop a simple and rapid technology for high efficiency heterologous protein production in suspension-cultured transgenic rice cells. Here, five keystone technology will be developed：（1）A whole new design of rice suspension cell expression cassette for multiple variable proteins production；（2）A simple and rapid transformation system；（3）An application of one effective rice peptide secret module；（4）the development of novel GFP production indicator which could simplify the monitor analysis operation during the developmental downstream process of this technology；（5）A scale-up immobilization fermentation process for efficient target proteins production. This proposal is the last part of a 3-year project by National science and technology program for agricultural biotechnology. We have great harvest in the last two years as: (1) the rice suspension cultured system has been successfully induced and established, (2) the analysis technique of GM-CSF by ELISA has been developed, (3) the high GM-CSF expression transgenic cell lines have been successfully selected and scaled up, (4) the laboratory-scale suspension and immobilization fermentation processes for efficient target proteins production have been investigated and compared (5) the optimum cultivation strategies for transgenic rice cell culture producing human serum have been investigated and modified. Following the previous two-year results, we will investigate the optimal production condition and operation model by the technology platform. The bioreactor study will be performed to produce the high-yield GM-CSF fermentation broth. The final purpose is to design and analyze an appropriate bioreactor for the continuous production of GM-CSF. Moreover, the equipment and cost should be reduced as low as possible. By combination and coherence of different high technology with high efficiency and low cost, we can predict the tremendous potential of this heterologous protein production technology platform.

Project ID:PD9706-0286
External Project ID:NSC97-2317-B182-001