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High throughput quantitative analysis of plant N-glycan using DNA sequencing equipment

첫 페이지 보기
  • 발행기관
    한국당과학회 바로가기
  • 간행물
    한국당과학회 학술대회 바로가기
  • 통권
    2008 Eastern Asian Glycoscience Symposium (2008.11)바로가기
  • 페이지
    pp.41-42
  • 저자
    Kyung Jin Lee, Jin-Hee Jung, Jung Mi Lee, Ohsuk Kwon, Hyun Ah Kang, Kisung Ko, Doo-Byoung Oh
  • 언어
    영어(ENG)
  • URL
    https://www.earticle.net/Article/A192309

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원문정보

초록

영어
Rapid and reliable quantitative plant N-glycan analysis is highly required to explore complex glycobiological research and develop plant expression systems for factory to produce therapeutic glycoproteins with human compatible glycans. We present here the high throughput quantitative analytical method for plant N-glycan based on DNA sequencing equipment. All steps including peptide N-glycosidase (PNGase) A treatment, glycan preparation and exoglycosidase digestion were optimized for high throughput applications with 96 well format procedures and automatic analysis on DNA sequencer. The glycans of horseradish peroxidase with plant-specific core α(1,3)- fucose can be discriminated by the comparison of glycan profiles obtained by PNGase A and F treatments. The peaks of glycans with (91%) and without (1.2%) α(1, 3)- fucose were easily quantified on a DNA sequencer analysis. In addition, both of them have been shown to harbor bisecting β(1,2)-xylose by simultaneous treatment of α (1,3)-mannosidase and β(1,2)-xylosidase. This technique was also applied to analyze N-glycan of plant-derived anti-rabies monoclonal antibody of which heavy chain was fused to C-terminal Lys-Asp-Glu-Leu (KDEL) sequences for the retention at endoplasmic reticulum. It revealed that most of its N-glycan (more than 80%) belongs to oligomannose type. The identities of glycans generated by DNA sequencer analysis were independently confirmed by mass spectrometry. These results suggest that the DNA sequencer- based method provides quantitative information for plant specific N-glycan analysis in a high throughput manner, which has not been achieved by glycan profiling based on mass spectrometry.

저자

  • Kyung Jin Lee [ Omics and Integration Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), Department of Biological Science, College of Natural Sciences, Wonkwang University ]
  • Jin-Hee Jung [ Omics and Integration Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB) ]
  • Jung Mi Lee [ Omics and Integration Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB) ]
  • Ohsuk Kwon [ Omics and Integration Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB) ]
  • Hyun Ah Kang [ Department of Life Science, College of Natural Science, Chung-Ang University ]
  • Kisung Ko [ Department of Biological Science, College of Natural Sciences, Wonkwang University ]
  • Doo-Byoung Oh [ Omics and Integration Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB) ]

참고문헌

자료제공 : 네이버학술정보

간행물 정보

발행기관

  • 발행기관명
    한국당과학회 [Korean Society for Glycoscience]
  • 설립연도
    2006
  • 분야
    의약학>약학
  • 소개
    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.

간행물

  • 간행물명
    한국당과학회 학술대회
  • 간기
    연간
  • 수록기간
    2006~2022
  • 십진분류
    KDC 517 DDC 614

이 권호 내 다른 논문 / 한국당과학회 학술대회 2008 Eastern Asian Glycoscience Symposium

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