Bioactivity and efficacy of therapeutic protein are dependent on structure-specific glycan attached to the therapeutic proteins. Therefore, precise glycomic characterization and monitoring are essential in the manufacture of therapeutic glycoproteins. However, the inherent structural diversity of glycosylation significantly hinders analysis. Thus, it is challenging to comprehensively characterize structure–specific glycans. We propose isomer-specific LC/MS and MS/MS screening as a method for rapid identification and structural elucidation of biopharmaceutical glycosylation. Biotherapeutic glycoproteins including mAbs, and EPOs were prepared and analyzed. After removing salts and detergents, N-glycans were enzymatically released and purified by graphitized carbon solid phase extraction. PGC chip nano-LC/Q-TOF provided chromatographic isomer separation profiling and structural characterization. Multiple structure isomers including antennary, polylactosamine, alpha galactose epitope, and linkage iosomers were identified within isomeric compositions. A database of accurate mass, RT, and diagnostic MS/MS fragments was developed and applied to identify and structurally characterize biopharmaceutical glycans. The approach is fast, does not need for labeling or derivatisation and is applicable for highly accurate identification of biotherapeutic glycosylation.
저자
Yi-Nae Jeon [ Graduate School of Analytical Science and Technology, Chungnam National University, Asia- Pacific Glycomics Reference Site, 99 Daehak-ro, Yuseong-gu, Daejeon, 34134, Republic of Korea ]
Myung Jin Oh [ Graduate School of Analytical Science and Technology, Chungnam National University, Asia- Pacific Glycomics Reference Site, 99 Daehak-ro, Yuseong-gu, Daejeon, 34134, Republic of Korea ]
Hyun Joo An [ Graduate School of Analytical Science and Technology, Chungnam National University, Asia- Pacific Glycomics Reference Site, 99 Daehak-ro, Yuseong-gu, Daejeon, 34134, Republic of 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.