나노 액체크로마토그래피-텐덤 질량분석기를 이용하여 N-당질화 위치 및N-당사슬 구조 규명을 위한 방법 (A Sensitive Method for Identification of N-Glycosylation Sites and the Structures of N-Glycans Using Nano-LC-MS/MS)

서지정보

· 발행기관 : 대한약학회
· 수록지 정보 : 약 학 회 지 / 57권 / 4호 / 250 ~ 257페이지
· 저자명 : 조영은, 김숙경, 백문창

초록

Biosimilars are important drugs in medicine and contain many glycosylated proteins. Thorough analysis of theglycosylated protein is a prerequisite for evaluation of biosimilar glycan drugs. A method to assess the diversity of N-glycosylationsites and N-glycans from biosimilar glycan drugs has been developed using two separate methods, LC-MS/MSand MALDI-TOF MS, respectively. Development of sensitive, accurate, and efficient methods for evaluation of glycoproteinsis still needed. In this study, analysis of both N-glycosylation sites and N-glycans of glycoprotein was performed using thesame LC-MS/MS with two different nano-LC columns, nano-C18 and nano-porous graphitized carbon (nano-PGC) columns.
N-glycosylated proteins, including RNAse B (one N-glycosylation site), Fetuin (three sites), and α-1 acid glycoprotein (foursites), were used, and small amounts of each protein were used for identification of N-glycosylation sites. In addition, highmannose N-glycans (one type of typical glycan structure), Mannose 5 and 9, eluted from RNAse B, were successfully identifiedusing nano-PGC-LC MS/MS analysis, and the abundance of each glycan from the glycoprotein was calculated. Thisstudy demonstrated an accurate and efficient method for determination of N-glycosylation sites and N-glycans of glycoproteinsbased on high sensitive LC-MS/MS using two different nano-columns; this method could be applied for evaluationof the quality of various biosimilar drugs containing N-glycosylation groups

영어초록

Biosimilars are important drugs in medicine and contain many glycosylated proteins. Thorough analysis of theglycosylated protein is a prerequisite for evaluation of biosimilar glycan drugs. A method to assess the diversity of N-glycosylationsites and N-glycans from biosimilar glycan drugs has been developed using two separate methods, LC-MS/MSand MALDI-TOF MS, respectively. Development of sensitive, accurate, and efficient methods for evaluation of glycoproteinsis still needed. In this study, analysis of both N-glycosylation sites and N-glycans of glycoprotein was performed using thesame LC-MS/MS with two different nano-LC columns, nano-C18 and nano-porous graphitized carbon (nano-PGC) columns.
N-glycosylated proteins, including RNAse B (one N-glycosylation site), Fetuin (three sites), and α-1 acid glycoprotein (foursites), were used, and small amounts of each protein were used for identification of N-glycosylation sites. In addition, highmannose N-glycans (one type of typical glycan structure), Mannose 5 and 9, eluted from RNAse B, were successfully identifiedusing nano-PGC-LC MS/MS analysis, and the abundance of each glycan from the glycoprotein was calculated. Thisstudy demonstrated an accurate and efficient method for determination of N-glycosylation sites and N-glycans of glycoproteinsbased on high sensitive LC-MS/MS using two different nano-columns; this method could be applied for evaluationof the quality of various biosimilar drugs containing N-glycosylation groups

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