Protein glycosylation with structural diversity is one of the most abundant post translational modifications. A wide range of functions for glycans has been described, from structural to regulatory roles in molecular and cellular level. Especially, sialic acid and its modifications are known to affect stability, efficacy, antigenicity, and half-life of biopharmaceuticals. Current glycosylation profiling is mainly focused on N-glycans due to availability of enzymes for their release. In contrast, there is no corresponding enzymes for O-glycan liberation. In general, O-glycans are liberated using alkaline β-elimination which conditions are highly destructive toward sialic acid and its modifications. For this reason, comprehensive O-glycosylation profiling method which preserves the sialic acid and its modifications is required to demonstrate in respect of quality, safety and efficacy of therapeutic glycoproteins. In this study, we developed a new analytical strategy for non-destructive O-glycosylation profiling of biopharmaceuticals using O-glycopeptides produced by non-specific protease. Erythropoietins (EPOs) were digested with pronase E for six hours, and then resulting glycopeptides were enriched using a solid phase extraction with porous graphitized carbon. The biopharmaceutical O-glycosylation profiling and structural characterization were conducted on nanoLC-chip/Q-TOF MS system. We successfully applied this method to analysis of batch-to-batch variation study of darbepoetin alfa, and O-glycosylation profiling of epoetin alfa and beta.
저자
Unyong Kim [ Asia Glycomics Reference Site, Daejeon, Korea, Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Korea ]
Myung Jin Oh [ Asia Glycomics Reference Site, Daejeon, Korea, Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Korea ]
Hyun Joo An [ Asia Glycomics Reference Site, Daejeon, Korea, Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Korea ]
본 학회는 화학, 생화학, 분자생물학, 미생물학, 식품공학, 의학, 약학, 유전공학 및 생물공학, 환경 및 기타 공업 등 전 분야의 탄수화물관련 이론과 기술을 연구 발전시키고 산학협동을 통해 이를 보급하여 국내 관련 산업의 발전 및 국민생활의 과학화에 기여하고자 하며, 이러한 목표와 비젼의 실현을 위해 회원들이 적극적인 참여와 활동을 전개하고자 한다.