title: An enzyme-linked immunosorbent assay for the measurement of Lewis blood-group alpha-(1----4)-fucosyltransferase activity.
abstract: An enzyme-linked immunosorbent assay (ELISA) has been developed for the Lewis blood-group associated alpha-(1----4)-fucosyltransferase activity. Microtiter plates coated with the bovine serum albumin conjugate of a synthetic beta-D-Galp-(1----3)-beta-D-GlcpNAc disaccharide are incubated with a fucosyltransferase preparation in the presence of guanosine 5'-diphosphofucose. The resulting immobilized Lewis-a active trisaccharide beta-D-Galp-(1----3)-[alpha-L-Fucp-(1----4)]-beta-D-GlcpNAc is then detected and quantitated using a monoclonal anti-Lewis-a antibody. Product formation detected in this manner is linear with time, proportional to enzyme activity, and reproducibly quantitated in the 50-400 fmol range.
authors.affiliation: Department of Food Science, University of Alberta, Edmonton, Canada.
authors.forename: M M
authors.initials: MM
authors.lastname: Palcic
authors.forename: R M
authors.initials: RM
authors.lastname: Ratcliffe
authors.forename: L R
authors.initials: LR
authors.lastname: Lamontagne
authors.forename: A H
authors.initials: AH
authors.lastname: Good
authors.forename: G
authors.initials: G
authors.lastname: Alton
authors.forename: O
authors.initials: O
authors.lastname: Hindsgaul
mesh_terms: D002240:Carbohydrate Sequence; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005647:Fucosyltransferases / Q000097:blood*; D006602:Hexosyltransferases / Q000097:blood*; D006801:Humans; D007983:Lewis Blood Group Antigens; D008969:Molecular Sequence Data
chemical_list: D007983:Lewis Blood Group Antigens; D005647:Fucosyltransferases; D006602:Hexosyltransferases; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase