Development of lectin-based lateral flow assay for fucosylated alpha-fetoprotein.

Abstract

Fucosylated alpha-fetoprotein (AFP-L3) is a more specific and sensitive biomarker for early diagnosis of hepatocellular carcinoma (HCC) than only the alpha-fetoprotein (AFP) level. Rapid and simple detection of AFP-L3 level greatly facilitates the early detection as well as the treatment of HCC, resulting in the reduction of mortality. Here, we developed a rapid and sensitive lateral flow assay (LFA) using lectin Lens culinaris agglutinin (LCA), which has a specific affinity to AFP-L3 fraction of AFP, as a biorecognition element for determination of the fucosylation of AFP. The assay is based on a sandwich format performed on a lateral flow test strip. LCA was immobilized on the membrane as a test line (T). Quantitative detection of AFP-L3 was achieved by measuring the green color intensity of captured gold nanoparticle conjugates on the T and control line (C) utilizing an in-house test strip reader. The calculated absorbance obtained by the green color intensity signals proportionally increased with AFP concentrations. The developed lectin-based LFA provided a detection limit of 0.8 ng/mL for AFP with a linear range between 1.5 and 160.0 ng/mL within an assay time of 10 min. Recoveries between 74.5% and 113.2% with relative standard deviations of 5.2%-8.7% for measuring spiked human serum were also achieved. The results reveal that the proposed assay offers a rapid, sensitive, and specific method, which is useful for development in point-of-care testing for early detection and treatment of HCC.