Novel method for quantifying alpha(1----3)-L-fucosyltransferase activity in serum

Area of Science:

• Biochemistry
• Immunology
• Clinical Chemistry

Background:

• Alpha(1----3)-L-fucosyltransferase enzyme activity is implicated in various biological processes.
• Quantifying this enzyme activity in human serum presents analytical challenges.

Purpose of the Study:

• To develop and validate a novel assay for quantifying alpha(1----3)-L-fucosyltransferase activity in human sera.
• To evaluate the diagnostic potential of this assay in distinguishing gastric disorders.

Main Methods:

• A sandwich-type immunoradiometric assay was developed using H type 2 trisaccharide-BSA conjugate.
• Guanylyl transferase (GDP)-fucose was used as a substrate, and the product was detected using specific antibodies.
• Assay validation included inter- and intra-assay variability, dilution linearity, and analytical recovery.

Main Results:

• The assay demonstrated high precision with inter- and intra-assay coefficients of variation (CVs) below 4%.
• Satisfactory results were obtained for dilution linearity and analytical recovery.
• Serum alpha(1----3)-L-fucosyltransferase activity was measured in healthy subjects and patients with benign and malignant gastric disorders.
• The assay detected higher enzyme activity in cancer patients compared to controls, especially in early stages, outperforming carcinoembryonic antigen (CEA).

Conclusions:

• The developed assay is a reliable and sensitive method for determining serum alpha(1----3)-L-fucosyltransferase activity.
• This assay shows promise as an early diagnostic marker for gastric cancer, detecting elevated enzyme levels.
• The findings suggest alpha(1----3)-L-fucosyltransferase activity is a valuable biomarker for early gastric cancer detection.