title: Enzyme-linked immunosorbent assays for the measurement of blood group A and B glycosyltransferase activities.
abstract: ELISA assays have been developed for alpha(1-3)N-acetylgalactosaminyltransferase (blood group A transferase) and alpha(1-3)galactosyltransferase (blood group B transferase) activities. In these assays, microtitre plates coated with the bovine serum albumin conjugate of a synthetic Fuc alpha 1-2Gal beta-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 Fuc alpha 1-2(GalNAc alpha 1-3)Gal beta-R-BSA or Fuc alpha 1-2(Gal alpha 1-3)Gal beta-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.
authors.affiliation: Department of Food Science, University of Alberta, Edmonton, Canada.
authors.forename: L M
authors.initials: LM
authors.lastname: Keshvara
authors.forename: E M
authors.initials: EM
authors.lastname: Newton
authors.forename: A H
authors.initials: AH
authors.lastname: Good
authors.forename: O
authors.initials: O
authors.lastname: Hindsgaul
authors.forename: M M
authors.initials: MM
authors.lastname: Palcic
mesh_terms: D002240:Carbohydrate Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D005700:Galactosyltransferases / Q000032:analysis*; D006801:Humans; D008969:Molecular Sequence Data; D017350:N-Acetylgalactosaminyltransferases / Q000032:analysis*
chemical_list: D005700:Galactosyltransferases; D017350:N-Acetylgalactosaminyltransferases; C003742:UDPgalactosamine-galactose acetylgalactosaminyltransferase; C023155:blood-group-substance alpha-D-galactosyltransferase