Effects of carbon sources and plant-derived peptone on hCTLA4Ig production in transgenic rice cell cultures

초록

Plant-derived peptones have been utilized to improve cell growth and productivity in plant cell cultures over 10 years. Concentration of carbon sources in culture media had to be optimized according to specific plant species and culture methods. Carbon and nitrogen sources have been known as major factors in plant cell cultures. Therefore, it was necessary to investigate the effects of carbon and nitrogen sources for the improvement of recombinant protein production. Supplementation of defined medium with plant-derived peptone was investigated to enhance cell growth and the production of human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) in transgenic rice cell suspension cultures. In addition, optimization of the concentrations of carbon and nitrogen sources is required. As a carbon source, 0 (control), 3, 5% (w/v) of sucrose was added in 100-mL Erlenmeyer flasks with 10 g/L plant-derived peptone. In case of adding 5% sucrose, the maximum dry cell weight and hCTLA4Ig production were higher than those of control culture. To examine the effect of nitrogen source, 0 (control), 10, 20, 40 g/L of plant-derived peptone was added in 100-mL Erlenmeyer flasks with 3% sucrose. Addition of 10 g/L of plant-derived peptone, the maximum dry cell weight was 1.9-fold higher than that of control and hCTLA4Ig production was 64.8 mg/L. hCTLA4Ig was not produced in control. Addition of plant-derived peptone could produce the hCTLA4Ig without media exchange. In conclusion, plant-derived peptone provided the enhancement of hCTLA4Ig production without media exchange. Determination of optimal ratio between carbon and nitrogen sources was found to be important to improve cell growth and hCTLA4Ig production in transgenic rice cultures.