PC7-1 : 2D-LC (HILIC & WAX) Analysis using Enzyme array

시험소개

목적 당단백질에 결합된 전체 N-glycan에 대한 Exoglycosidase 처리에 의한 전체 당구조의 분석으로 HILIC 과 WAX 분석을 통해 목적 단백질에 결합된 Oligosaccharide의 함량을 분석하는 방법이다.

시험 대상

  • Biosimilar
  • Biobetter
  • Antibody
  • Antibody-drug conjugates
  • Recombinant protein drug
  • Toxin
  • vaccine

시험 방법

  • N-Glycan Release by
    PNGase-F
  • 2-AB derivatization
  • HILIC-UPLC
    WAX-UPLC
  • Exoglycosydase
  • HILIC-UPLC
    WAX-UPLC

시험결과

좌우로 스크롤하여 내용을 확인하세요.
Peaka GUb %Totalc Exoglycosidase digestionsd Chargee Namef
F S SG SGN
14 15.04 7.4 -4 FA4G4Lac2S4
14-1 14.93 8.3 A4G4Lac2S4
14-2 12.54 16.2 FA4G4Lac2
14-3 9.28 15.3 FA4G2N2
14-4 4.80 100.0 FM3
  • aPeak number in Figures.
  • bGlucose unit. Average GU is used in the case of is omer peaks.
  • cAmount of gly can expressed as a percentage of the total glycans measured by HILIC, which is coverted to a relative value in whole identified peaks.
  • dF, α(1-2,3,4,6)-Fucosidase; S, Sialidase A;G,β(1-4,6)-Galactosidase;N,β-N-Acetylhexosaminidase.
  • edetermined by WAX.
  • fF, Fucose; A, antenna; G, galactose; Lac, lactosamine; S, sialic acid; M, mannose; N, N-acetylglucosamine.
    Each number indicates the number of monosaccharide or disaccharide in the corresponding oligosaccharide.
    So, FA4G4Lac2S4 represents core fucosylated, tetra-sialylated, galactosylated tetraantennary glycan with two lactosamine.
    Symbolic representation proposed by Harvey et al (Proteomics 2009. 9:3796) is used for the report.