Application of a Novel One-Side Cell Quartz Crystal Microbalance Immunosensor in the Determination of Alpha-Fetoprotein from Human Serum

新型单侧细胞石英晶体微天平免疫传感器在人血清甲胎蛋白测定中的应用

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作者:Yan Chen, Huashan Shi, Bo Mu

Abstract

The rapid and accurate detection of alpha-fetoprotein (AFP) levels is of great significance for the diagnosis and later treatment evaluation of liver cancer. In this study, a novel integrated quartz crystal microbalance (QCM) immunosensor based on the design to contact liquids on one side only was developed for the label-free detection of AFP. Anti-AFP mouse monoclonal antibodies were immobilized onto the upper electrode surface of the pre-treatment crystals using the staphylococcus aureus protein A. The AFP antigens in human serum were captured by specific surface-coated antibodies, and testing was carried out by monitoring the corresponding changes in frequency. The concentration range of the antigen detected was 13-760 ng/mL. The frequency characteristics of the process of antibody immobilization were investigated in detail, and high reproducibility of AFP antibody immobilization was achieved (standard deviation (SD) = 2.2 Hz). The developed QCM measurement system demonstrated a short test time (only 30 min), good reproducibility (the biological activity can still maintain more than 90% of the first test till it is reused five times), and accuracy as good as the one achieved with the radioimmunoassay (the maximum relative deviation = 4.8%). The designed QCM test system can easily and quickly detect AFP concentrations up to 760 ng/mL, indicating that the developed QCM assay is likely to lead to an alternative approach in large-scale screening for liver cancer in the near future.

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