Supplementation of carbon sources and plant-derived peptone for the enhanced production of hCTLA4Ig in transgenic rice cell cultures

초록

For over a century, peptones have been used as the main nutritive components in cell culture media. Peptones contain peptides, amino acids, minerals, and other micronutrients that may serve as growth factors and help mammalian cell metabolic processes, thereby promoting cell proliferation and production of recombinant proteins. In terms of regulatory approval of produced biopharmaceuticals, plant-derived peptones are preferred rather than animal-derived because of the absence of animal-derived additives and impurities. Plant-derived peptones have been also utilized to improve cell growth and productivity in plant cell cultures over 10 years. To enhance the volumetric productivity, optimization of concentration ratio between plant-derived peptone and sucrose is important because 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 performed 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 1% plant-derived peptone. In case of adding 5% sucrose, maximum dry cell weight and hCTLA4Ig production were higher than those of control culture. To examine the effects of nitrogen source, 0 (control), 10, 20, 40 g/L of plant-derived peptone was added in 100-mL Erlenmeyer flasks with 3% sucrose.