Biotechnology and Bioprocess Engineering:BBE

  • 제5권2호
  • /
  • Pages.136-140
  • /
  • 2000
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  • 1226-8372(pISSN)
  • /
  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Quantitative Analysis of Oligosaccharide Structure of Glycoproteins

  • Chang, Kern-Hee (Department Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1, kusung dong, Young gu, Taejon 305-701) ;
  • Tamao Endo (Department of Glycobiology, Tokyo Metropolitan Institute of Gerontology, 35-2 Sakae-cho, Itabashi-ku, Tokoy 173-0015,Japen) ;
  • Kim, Jung-How (Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1, kusung dong, Young gu, Taejon 305-701)
  • 발행 : 2000.04.01
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초록

A sensitive and quantiative method for the structural analysis of oligosaccharide was established for the glycoform analysis of glconproproteins. Inthis study, n-linked oligosaccharides of human IgG and bovine transferin were analyzed for the evaluation of the methydrate moiety ofthe method. Chrbohydrate moiety of glycoprotein was relased by hydrazinolysis and purified by paper chromatography. The oligosaccharides were labeled with a fluorescent bye, 2-aminobenzamide, for the enhancement of detection sensitivity. sialylated (acidic) oligosaccharides were separated from neutral oligosaccharide by employing a strong anion-exchange column(MonoQ) followed by the treatment with sialidase. Enzymatically desiayated fractions and neutral fractions of oligosaccharides were applied to normal-phase HPLC to resolve the peaks according to glucose unit (GU). The structure of separated molecules was further determined by sequential digestion with exoglycosidases. As a result, disialylated biantennary complextype oligosaccharide was found to be a major sugar chain in bovine transferrin (63%). In human IgG, core fucosylated asialobiantennary complex oligosaccharides were dominant. These results coincided well with reported results.

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참고문헌

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