Utilization of Kifunensine for the Production of Acid α-glucosidase with High-mannose Type N-glycans in Transgenic Rice Cell Cultures

초록

Pompe disease is one of lysosomal storage diseases caused by dysfunction of acid α-glucosidase (GAA).Enzyme replacement therapy is known to be the most effective for the treatment of Pompe disease through mannose 6-phosphate (M6P) pathway. Therefore, production of GAA including high-mannose type N-glycan is important to make M6P structure. Kifunensine is a small molecule inhibitor of class I α-mannosidase in endoplasmic reticulum and Golgi apparatus [1,2]. Inhibition of mannosidase could reduce plant-specific glycan residues which have potential immunogenicity to humans. In this study, three different concentrations of kifunensine (5, 10, 20 μM) were added to transgenic rice cell culture medium. toxic effects in both cell growth and viability were not shown during cell cultures. Composition of complex type N-glycans was reduced below 10% and Man 8-9 glycans lacking core-α1,3-fucose, core-β1,2-xylose was increased over 50%. In conclusion, it was found that kifunensine could be used as an N-glycan regulator during in vitro cell cultures for the production of therapeutic proteins with humanized N-glycans.