Novel method for quantifying alpha(1----3)-L-fucosyltransferase activity in serum.
A novel method has been developed to quantify alpha(1----3)-L-fucosyltransferase activity in human sera by applying a sandwich-type immunoradiometric assay. H type 2 trisaccharide (6Fuc alpha 1----2Gal beta 1----4GlcNAc) covalently attached to bovine serum albumin (BSA) was used as an acceptor and incubated with serum samples in the presence of guanosine diphosphate-fucose. The resulting product, Y tetrasaccharide (Fuc alpha 1----2Gal beta 1----4[Fuc alpha 1----3] GlcNAc beta-BSA), was detected by a sequential use of anti-BSA antibody-coated bead and 125I-labeled anti-Y antibody. Inter- and intra-assay CVs for alpha(1----3)-L-fucosyltransferase were both less than 4%, and the results of the dilution linearity and analytical recovery studies were satisfactory. Using the present assay method, we measured alpha(1----3)-L-fucosyltransferase in serum from patients with benign and malignant gastric disorders and in healthy subjects. The detection rate of alpha(1----3)-L-fucosyltransferase for cancer was apparently higher than that of carcinoembryonic antigen measured in the same samples, particularly in the early clinical stage; indeed, no correlation was observed between the concentrations of the two potential markers. The results indicate that the present assay method seems to be excellent for the determination of serum alpha(1----3)-L-fucosyltransferase activity and useful for the detection of cancer-associated increases of the enzyme activity at the early stage of gastric cancer.
Tachikawa
T
T
Diagnostic Division, Otsuka Pharmaceutical Co., Ltd., Tokushima, Japan.
Yazawa
S
S
Asao
T
T
Shin
S
S
Yanaihara
N
N
D002240:Carbohydrate Sequence; D002272:Carcinoembryonic Antigen / Q000378:metabolism; D005647:Fucosyltransferases / Q000097:blood*; D006801:Humans; D015592:Immunoradiometric Assay / Q000706:statistics & numerical data; D008969:Molecular Sequence Data; D012710:Serum Albumin, Bovine; D013274:Stomach Neoplasms / Q000201:enzymology; D014312:Trisaccharides / Q000737:chemistry / Q000378:metabolism
D002272:Carcinoembryonic Antigen; D014312:Trisaccharides; D012710:Serum Albumin, Bovine; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase