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Enzyme-linked immunosorbent assays for the measurement of blood group A and B glycosyltransferase activities

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Abstract

ELISA assays have been developed for α(1–3)N-acetylgalactosaminyltransferase (blood group A transferase) and α(1–3)galactosyltransferase (blood group B transferase) activities. In these assays, microtitre plates coated with the bovine serum albumin conjugate of a synthetic Fucα1–2Galβ-R acceptor substrate are incubated with the appropriate nucleotide donor (UDP-GalNAc or UDP-Gal) and human serum as the enzyme source. The resulting trisaccharide products Fucα1–2(GalNAcα1–3)Galβ-R-BSA or Fucα1–2(Galα1–3)Galβ-R-BSA are detected and quantified with monoclonal antibodies selected not to cross-react with the substrate structure. With less than a microliter of human serum, product formation is proportional to enzyme concentration and to time of incubation of up to 90 min.

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Authors and Affiliations

  1. Department of Food Science, University of Alberta, T6G 2P5, Edmonton, Alberta, Canada

    Lakhu M. Keshvara, Elizabeth M. Newton & Monica M. Palcic

  2. Department of Chemistry, University of Alberta, T6G 2G2, Edmonton, Alberta, Canada

    Lakhu M. Keshvara, Elizabeth M. Newton & Ole Hindsgaul

  3. Chembiomed Ltd., Edmonton Research and Development Park, Station F, P.O. Box 8050, T6H 4N9, Edmonton, Alberta, Canada

    A. Heather Good

Authors
  1. Lakhu M. Keshvara
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  2. Elizabeth M. Newton
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  3. A. Heather Good
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  4. Ole Hindsgaul
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  5. Monica M. Palcic
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Keshvara, L.M., Newton, E.M., Good, A.H. et al. Enzyme-linked immunosorbent assays for the measurement of blood group A and B glycosyltransferase activities. Glycoconjugate J 9, 16–20 (1992). https://doi.org/10.1007/BF00731173

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  • DOI: https://doi.org/10.1007/BF00731173

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