Abstract
The 8-methoxycarbonyloctyl glycosides of GlcNAcβ, Galβ1-4Glcβ, Fucα1-2Fucα1-3GalNacβ and Fucα1-2Galβ1-3[Fucα1-4]GlcNacβ were converted to primary amines by reaction with neat ethylenediamine and then coupled to bovine serum albumin (BSA) using diethyl squarate as the connector. The average degree of incorporation of the sugar onto the protein, as well as the molecular weight distribution, could be conveniently determined using matrix assisted laser desorption ionization/time of flight (MALDI-TOF) mass spectrometry thus avoiding cumbersome structure-dependent colour-tests or analysis of cleaved ligand. The present coupling method has the advantages of proceeding under very mild conditions, yielding controlled incorporation values and can reliably be used for the coupling of very small amounts (mg) of oligosaccharide.
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References
Stowel CP, Lee YC (1980)Adv Carboydr Chem Biochem 37: 225–81.
Varki A (1993)Glycobiology 3: 97–130.
Hakomori S-I (1991)Pure Appl Chem 63: 473–82.
Lee YC, Lee RT, eds. (1994) Neoglycoconjugates,Methods Enzymol 242.
Magnusson G, Chernyak AY, Kihlberg J, Kononov LO (1994) InNeoglycoconjugates: Preparation and Application (Lee YC, Lee RT, eds), pp. 53–143, San Diego: Academic Press.
Tietze L, Arlt M, Beller M, Glusenkamp KH, Jahde E, Rajewsky MF (1991)Chem Ber 124: 1215–19.
Lemieux RU, Bundle DR, Baker DA (1975)J Am Chem Soc 97: 4076–83.
Pinto BM, Bundle DR (1983)Carbohydr Res 124: 313–18.
Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956)Anal Chem 28: 350–56.
Hindsgaul O, Norberg T, LePendu J, Lemieux RU (1982)Carbohydr Res 190: 109–42.
Banoub J, Bundle DR (1979)Can J Chem 57: 2085–90.
Kamath VP, Hindsgaul O (1995)Carbohydr Res (in press).
Spohr U, Lemicux RU (1988)Carbohydr Res 174: 211–37.
Zhang Y, Le X, Dovichi NJ, Compston CA, Palcic MM, Diedrich P, Hindsgaul O (1995)Anal Biochem 227: 368–76.
Hällgren C, Hindsgaul O (1995)J Carbohydr Chem 14: 453–64.
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This paper is dedicated to Sen-itiroh Hakomori on the occasion of his 65th birthday
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Kamath, V.P., Diedrich, P. & Hindsgaul, O. Use of diethyl squarate for the coupling of oligosaccharide amines to carrier proteins and characterization of the resulting neoglycoproteins by MALDI-TOF mass spectrometry. Glycoconjugate J 13, 315–319 (1996). https://doi.org/10.1007/BF00731506
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DOI: https://doi.org/10.1007/BF00731506