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Glycoconjugate Journal

, Volume 25, Issue 1, pp 11–14 | Cite as

2-Aminopyridine—a label for bridging of oligosaccharides HPLC profiling and glycoarray printing

  • N. V. Shilova
  • O. E. Galanina
  • A. Yu. Rubina
  • V. I. Butvilovskaya
  • M. E. Huflejt
  • J. Chambers
  • A. Roucoux
  • N. V. BovinEmail author
Article

Abstract

2-Aminopyridine derivatives of oligosaccharides (OS-AP) were printed onto microchips by two different ways. The first method is based on direct covalent insertion of OS-AP in polyacrylamide gel 3D chip. The second method is based on conversion of OS-AP into more reactive OS-aminoalditol followed by covalent printing onto NHS-activated glass slides. This approach extends the range of saccharides suitable for covalent printing due to availability of commercial OS-AP and easy high-performance liquid chromatography separation of glycoprotein N-chains in form of AP derivatives.

Keywords

2-aminopyridine Fluorescent label Glycoconjugates Glycochip Oligosaccharides 

Abbreviations

AP

2-aminopyridine

BSA

bovine serum albumin

4-DMP

4-dimethylaminopyridine

DMSO

dimethyl sulfoxide

Fmoc-Gly-SI

N-(9-fluorenylmethoxycarbonyl)-glycine, succinimide ester

Ig

immunoglobulin

YDS

(Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1)2-3,6Manβ1-4GlcNAcβ1-4GlcNAc

iM8

iso-maltooctaose, (Glcβ1-6)8

Mel

melibiose, Galα1-6Glc

NP

p-nitrophenol

OS

oligosaccharide

NHS

N-hydroxisuccinimide

PBS

phosphate-buffered saline

pNPA

poly(4-nitrophenyl acrylate)

Et3N

triethylamine

9OS

(Galβ1-4GlcNAcβ1-2Manα1)2-3,6Manβ1-4GlcNAcβ1-4GlcNAc

Notes

Acknowledgements

The authors are thankful to P. Obukhova for isolation of antibodies. This work was supported by NIH grants 1 R01 GM060938-06 and 2 U54 GM062116-06 and the Molecular and Cell Biology Program, RAS.

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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • N. V. Shilova
    • 1
  • O. E. Galanina
    • 1
  • A. Yu. Rubina
    • 2
  • V. I. Butvilovskaya
    • 2
  • M. E. Huflejt
    • 3
  • J. Chambers
    • 3
  • A. Roucoux
    • 4
  • N. V. Bovin
    • 1
    Email author
  1. 1.Shemyakin & Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Engelhardt Institute of Molecular BiologyRussian Academy of SciencesMoscowRussia
  3. 3.Cellexicon, Inc.La JollaUSA
  4. 4.National School of Chemistry of RennesRennesFrance

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