Simultaneous Fluorescent Labelling and Biotinylation of Oligosaccharides: A Versatile Approach to the Analysis of Oligosaccharide Structure and Function

  • Derek K. Toomre
  • Ajit Varki
Part of the BioMethods book series (BIOMETHODS)


Oligosaccharides participate in a variety of important biological functions, including interactions with specific receptors (1–4). A prerequisite to the discovery and exploration of such interactions is the comprehensive structural analysis of homogeneous oligosaccharide species. The purification of oligosaccharides from complex mixtures requires diverse techniques, as well as sensitive and specific detection. One way to achieve sensitive detection is to attach a chromophore to the reducing sugar by reductive amination (5, 6) (see Chapters 7,11,15,18 in this volume for other examples of such techniques). Fluorescent chromophores (in contrast to light-absorbing chromophores) can often permit detection in the low picomole range for high-pressure liquid chromatography (HPLC) and polyacrylamide gel electrophoresis (PAGE) techniques, and in the femtomole range in capillary electrophoresis. Furthermore, hydrophobic or anionic properties of the chromophoric groups can be exploited to improve oligosaccharide fractionation by either electrophoresis or HPLC (5–8). Often several multidimensional chromatographic techniques, based on different physical properties, are required to adequately resolve complex mixtures. Several examples have been described of “two-dimensional mapping” with fluorescent pyridylamino (PA)-coupled oligosaccharides (5, 9). More recently, 2-aminobenzamide has been employed as an alternate fluorescent tag which gives both nonselective and highly efficient coupling to oligosaccharides (10).


Ammonium Formate Reductive Amination Pyridine Acetate Fast Atom Bombard Troubleshooting Guide 
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  1. 1.
    Lee YC (1992) FASEB J. 6: 3193.PubMedGoogle Scholar
  2. 2.
    Kornfeld S (1992) Annu. Rev. Biochem. 61: 307.PubMedCrossRefGoogle Scholar
  3. 3.
    Bevilacqua MP and Nelson RM (1993) J. Clin. Invest. 91: 379.PubMedCrossRefGoogle Scholar
  4. 4.
    Varki A (1993) Glycobiology 3: 97.PubMedCrossRefGoogle Scholar
  5. 5.
    Hase S (1994) Methods Enzymol. 230: 225.PubMedCrossRefGoogle Scholar
  6. 6.
    Lee KB, Al-Hakim A, Loganathan D and Linhardt RJ (1991) Carbohydr. Res. 214: 155.PubMedCrossRefGoogle Scholar
  7. 7.
    Stack RJ and Sullivan MT (1992) Glycobiology 2: 85.PubMedCrossRefGoogle Scholar
  8. 8.
    Jackson P (1991) Anal. Biochem. 196: 238.PubMedCrossRefGoogle Scholar
  9. 9.
    Tomiya N, Lee YC, YoshidaT, WadaY, Awaya J, Kurono M and Takahashi N (1991) Anal. Biochem. 193: 90.PubMedCrossRefGoogle Scholar
  10. 10.
    Shao MC, Sokolik CW and Wold F (1994) Noncovalent Neoglycoproteins. In: YC Lee and RT Lee (eds) Neoglycoconjugates: Preparation and Applications, p. 225, Academic Press, San Diego.Google Scholar
  11. 11.
    Shao MC, Chen LM and Wold F (1990) Methods Enzymol. 184: 653.PubMedCrossRefGoogle Scholar
  12. 12.
    Hase S (1992) J. Biochem. (Tokyo) 112: 266.Google Scholar
  13. 13.
    Rothenberg BE, Hayes BK, Toomre D, Manzi AE and Varki A (1993) Proc. Natl. Acad. Sci. USA 90: 1 1939.Google Scholar
  14. 14.
    Toomre DK and Varki A (1994) Glycobiology 4: 653.PubMedCrossRefGoogle Scholar
  15. 15.
    Shao MC (1992) Anal. Biochem. 205: 77.PubMedCrossRefGoogle Scholar
  16. 16.
    Wilchek M and Bayer EA(1988) Anal. Biochem. 171: 1.Google Scholar
  17. 17.
    Honda S and Suzuki S (1984) Anal. Biochem. 142: 167.PubMedCrossRefGoogle Scholar
  18. 18.
    Savage D, Mattson G, Desai S, Nielander G, Morgansen S and Conklin E (eds), Avidin-Biotin Chemistry: A Handbook (1992) Pierce Chemical Co., Rock- ford, IL.Google Scholar
  19. 19.
    Green NM (1965) Biochem. J. 94: 23c.PubMedGoogle Scholar
  20. 20.
    Bigge JC, Patel TP, Bruce JA, Goulding PN, Charles SM and Parekh R (1995) Anal. Biochem. 230: 229.PubMedCrossRefGoogle Scholar

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© Birkhäuser Verlag Basel 1997

Authors and Affiliations

  • Derek K. Toomre
  • Ajit Varki

There are no affiliations available

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