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  • 02월 26일 17시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능

제115회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Analytical Platform Employing Graphitized Carbon Liquid Chromatography-Mass Spectrometry for Glycomic Characterization of Biotherapeutic Glycoproteins

등록일
2015년 2월 23일 16시 49분 32초
접수번호
2046
발표코드
ANAL.O-46 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
발표시간
금 11시 : 30분
발표형식
구두발표
발표분야
분석화학 - Oral Presentation of Young Analytical Chemists
저자 및
공동저자
서나리, 안현주*
충남대학교 분석과학기술대학원, Korea
Monoclonal antibodies (mAbs) represent one of the largest classes of drugs in development due to their great efficacy for treatment of diseases such as cancers and rheumatic therapies. It has been known that glycosylation of protein therapeutics can radically affect their biological activity, plasma half-life, immunogenicity, and stability. Therefore, analysis of glycosylation is crucial for proper characterization of biotherapeutics in order to ensure the quality, safety, and consistency of the product. However, glycan analysis still remains analytical challenge due to its inherent microheterogeneity and diversity including the large number of isomeric structure. Liquid chromatography coupled with mass spectrometry (LC/MS) has become a key technique for rapid glycan profiling with high sensitivity and structure elucidation using tandem MS (LC/MS/MS). LC separation is essential for glycan isomer analysis because MS alone does not distinguish isomeric glycans with same mass. Hydrophilic interaction chromatography (HILIC) and porous graphitized carbon (PGC) combined with LC/MS are widely used for glycan separation. In this study, we have evaluated the chromatographic performance of HILIC and PGC including isomer separation, sensitivity, and quantitation for native and 2-AB labeled N-glycans, respectively. All peaks eluted from LC were further assigned by triple quad (QQQ) MS. We found that higher sensitivity was obtained from 2AB labeled glycans separated by HILIC column while this approach requires additional sample steps including glycan labeling and desalting. On the other hand PGC is a powerful tool in separating isomers of native glycans including neutral and acidic species with minimum sample preparation. Our analytical platform can be used to provide a reference for the column choice and optimized separation conditions for glycan analysis of biopharmaceutical glycoproteins.

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