An enzyme-linked immunosorbent assay for the measurement of Lewis blood-group alpha-(1----4)-fucosyltransferase activity.
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.
Palcic
M M
MM
Department of Food Science, University of Alberta, Edmonton, Canada.
Ratcliffe
R M
RM
Lamontagne
L R
LR
Good
A H
AH
Alton
G
G
Hindsgaul
O
O
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
D007983:Lewis Blood Group Antigens; D005647:Fucosyltransferases; D006602:Hexosyltransferases; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase