Utilization of Select Peptone to Reduce hCTLA4Ig Degradation in Transgenic Rice Cell Cultures

초록

Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced in transgenic rice cell cultures using RAmy3D promoter which was induced by sugar starvation. In sugar-depleted medium, cell viability was rapidly decreased and cell lysis was also occurred by shear stress. As a result, degradation of target protein could be observed. Plant-based peptone as an organic nitrogen source was used to improve cell viability for effective production of recombinant protein (1). In was demonstrated that peptone, consisted of polypeptides, could be an alternative substrate for protease and reduce the degradation of protein. In this study, to investigate the effects of select peptone on cell viability and protease activity, hCTLA4Ig spiking test was performed in conditioned media with select peptone. The stability of hCTLA4Ig was confirmed by Western blot analysis. In conclusion, supplementation of select peptone was found to reduce the protein degradation and improve the production of the target protein in transgenic rice suspension cell cultures.