기술이전

 

당단백질내 존재하는 알파갈락토스의 선택적 분석법

관리번호
RS05202011170002
첨부파일
개발년도
2020
저자정보
유종신
참고문헌
Anal. Chem. 2020, 92, 13144−13154
관련장비
하이브리드 FT-ETD 질량분석기
응용분야
당단백질 의약품내 알파갈락토스의 선택적 분석법
연구분야
BT
분석/적용분야
BT
기술분야
BT
분석/공동연구지원
당단백질내 존재하는 알파갈락토스의 분석지원
요약
We describe a method to identify α-galactosyl N-glycopeptides in mice glycoproteins using liquid chromatography with tandem mass spectrometry with higher-energy collisional dissociation (HCD).
키워드
α‑Galactosyl Epitopes, N‑Glycoproteins, Fragment Ions, Higher-Energy Collisional Dissociation
연구개발성과
혈액내 알파갈락토스형 당단백질의 분석법
내 용
The α-galactosyl epitope is a terminal N-glycan moiety of glycoproteins found in mammals except in humans, and
thus, it is recognized as an antigen that provokes an immunogenic response in humans. Accordingly, it is necessary to analyze the α-galactosyl structure in biopharmaceuticals or organ transplants. Due to an identical glycan composition and molecular mass between
α-galactosyl N-glycans and hybrid/high-mannose-type N-glycans, it is challenging to characterize α-galactosyl epitopes in Nglycoproteins using mass spectrometry. Here, we describe a method to identify α-galactosyl N-glycopeptides in mice glycoproteins
using liquid chromatography with tandem mass spectrometry with higher-energy collisional dissociation (HCD). The first measure
was an absence of [YHM] ion peaks in the HCD spectra, which was exclusively observed in hybrid and/or high-mannose-type Nglycopeptides. The second complementary criterion was the ratio of an m/z 528.19 (Hex2HexNAc1) ion to m/z 366.14
(Hex1HexNAc1) ion (Im/z528/Im/z366). The measure of [Im/z528/Im/z366 > 0.3] enabled a clear-cut determination of α-galactosyl Nglycopeptides with high accuracy. In Ggta1 knockout mice, we could not find any α-galactosyl N-glycoproteins identified in WT
mice plasma. Using this method, we could screen for α-galactosyl N-glycoproteins from mice spleen, lungs, and plasma samples in a
highly sensitive and specific manner. Conclusively, we suggest that this method will provide a robust analytical tool for determination
of α-galactosyl epitopes in pharmaceuticals and complex biological samples.