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


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.


2-aminopyridine Fluorescent label Glycoconjugates Glycochip Oligosaccharides 





bovine serum albumin




dimethyl sulfoxide


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






iso-maltooctaose, (Glcβ1-6)8


melibiose, Galα1-6Glc








phosphate-buffered saline


poly(4-nitrophenyl acrylate)







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.


  1. 1.
    Blixt, O., Head, S., Mondala, T., Scanlan, C., Huflejt, M.E., Alvarez, R., Bryan, M.C., Fazio, F., Calarese, D., Stevens, J., Razi, N., Stevens, D.J., Skehel, J.J., van Die, I., Burton, D.R., Wilson, I.A., Cummings, R., Bovin, N., Wong, C.H., Paulson, J.C.: Printed covalent glycan array for ligand profiling of diverse glycan binding proteins. Proc. Nat. Acad. Sci. USA. 101, 17033–17038 (2004)PubMedCrossRefGoogle Scholar
  2. 2.
    Dyukova, V.I., Shilova, N.V., Galanina, O.E., Rubina, A.Yu., Bovin, N.V.: Design of carbohydrate multiarrays. BBA 1760, 603–609 (2006)PubMedGoogle Scholar
  3. 3.
    Xia, B., Kawar, Z.S., Ju, T., Alvarez, R.A., Sachdev, G.P., Cummings, R.D.: Versatile fluorescent derivatization of glycans for glycomic analysis. Nature Meth. 2, 845–850 (2005)CrossRefGoogle Scholar
  4. 4.
    Shilova, N.V., Bovin, N.V.: Fluorescent labels for the analysis of mono- and oligosaccharides. Rus. J. Bioorgan. Chem. 29, 309–324 (2003)CrossRefGoogle Scholar
  5. 5.
    Takahashi, N., Wada, Y., Awaya, J., Kurono, M., Tomiya, N.: Two-dimensional elution map of GalNAc-containing N-linked oligosaccharides. Anal. Biochem. 208, 96–109 (1993)PubMedCrossRefGoogle Scholar
  6. 6.
    Takahashi, N.: Three-dimensional mapping of N-linked oligosaccharides using anion-exchange, hydrophobic and hydrophilic interaction modes of high-performance liquid chromatography. J. Chromatogr. A. 720, 217–225 (1996)PubMedCrossRefGoogle Scholar
  7. 7.
    Mevellec, V., Roucoux, A.: Nanoheterogeneous catalytic hydrogenation of N-, O- or S- heteroaromatic compounds by re-usable aqueous colloidal suspensions of rhodium(0). Inorg. Chim. Acta 357, 3099–3103 (2005)CrossRefGoogle Scholar
  8. 8.
    Bovin, N.V., Korchagina, E.Yu., Zemlyanukhina, T.V., Byramova, N.E., Galanina, O.E., Zemlyakov, A.E., Ivanov, A.E., Zubov, V.P., Mochalova, L.V.: Synthesis of polymeric neoglycoconjugates based on N-substituted polyacrylamide. Glycoconj. J. 10, 142–151 (1993)PubMedCrossRefGoogle Scholar
  9. 9.
    Bayer, E.A., Wilchek, M.: The use of a homologous series of affinity labeling reagents in the study of the biotin transport system in yeast cells. FEBS Lett. 60, 309–312 (1975)PubMedCrossRefGoogle Scholar
  10. 10.
    Khraltsova, L.S., Sablina, M.A., Melikhova, T.D., Joziasse, D.H., Kaltner, H., Gabius, H.J., Bovin, N.V.: An enzyme-linked lectin assay for α1,3-galactosyltransferase. Anal. Biochem. 280, 250–257 (2000)PubMedCrossRefGoogle Scholar
  11. 11.
    Hase, S., Hara, S., Matsushima, Y.: Tagging of sugars with a fluorescent compound, 2-aminopyridine. J. Biochem. 85, 217–220 (1979)PubMedGoogle Scholar
  12. 12.
    Nashabeh, W., Rassi, Z.E.: Capillary zone electrophoresis of linear and branched oligosaccharides. J. Chromatogr. 600, 279–287 (1992)PubMedCrossRefGoogle Scholar
  13. 13.
    Pukhalsky, A.L., Shmarina, G.V., Kalashnikova, E.A., Shiyan, S.D., Kokarovtseva, S.N., Pukhalskaya, D.A., Bovin, N.V.: Immunomodulating and antiinflammatory activity of semisynthetic analog of α1-acid glycoprotein. Bull. Exp. Biol. Med. 129, 567–570 (2000)CrossRefGoogle Scholar
  14. 14.
    Takahashi, C., Nakakita, S., Hase, S.: Conversion of pyridylamino sugar chains to corresponding reducing sugar chains. J. Biochem. 134, 51–55 (2003)PubMedCrossRefGoogle Scholar
  15. 15.
    Shiyan, S.D., Nasonov, V.V., Markin, V.A., Belyanchikov, I.M.: HPLC of N-(4-methylcoumarine-7-yl)glycamines in structural studies of N-linked glycoprotein carbohydrate chains. Bioorgan. Khim. 14, 1197–1207 (1988)Google Scholar
  16. 16.
    Dyukova, V.I., Dementieva, E.I., Zubtsov, D.A., Galanina, O.E., Bovin, N.V., Rubina, A.Yu.: Hydrogel glycan microarrays. Anal. Biochem. 347, 94–105 (2005)PubMedCrossRefGoogle Scholar
  17. 17.
    Korchagina, E.Yu., Bovin, N.V.: Synthesis of spacered trisaccharides with blood group specificities A and B, their fragments and structural analogs. Bioorgan. Khim. 18, 283–298 (1992)Google Scholar

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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