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Enzyme-linked immunosorbent assays for the measurement of blood group A and B glycosyltransferase activities |
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| Authors: | Lakhu M Keshvara Elizabeth M Newton A Heather Good Ole Hindsgaul Monica M Palcic |
| Institution: | (1) Department of Food Science, University of Alberta, T6G 2P5 Edmonton, Alberta, Canada;(2) Department of Chemistry, University of Alberta, T6G 2G2 Edmonton, Alberta, Canada;(3) Chembiomed Ltd., Edmonton Research and Development Park, Station F, P.O. Box 8050, T6H 4N9 Edmonton, Alberta, Canada |
| Abstract: | ELISA assays have been developed for  (1–3)N-acetylgalactosaminyltransferase (blood group A transferase) and (1–3)galactosyltransferase (blood group B transferase) activities. In these assays, microtitre plates coated with the bovine serum albumin conjugate of a synthetic Fuc1–2Gal -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 Fuc1–2(GalNAc1–3)Gal-R-BSA or Fuc1–2(Gal1–3)Gal-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. |
| Keywords: | ELISA blood group AN-acetylgalactosaminyltransferase blood group B galactosyltransferase glycosyltransferase assays |
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