Abstract
Background
Alpha-fetoprotein (AFP) has been used as a biomarker for early diagnosis of hepatocellular carcinoma (HCC) for a long time. Aptamer is a type of material that is receiving attention as a new alternative in the diagnosis and treatment of diseases.
Objective
We applied aptamer to AFP detection and utilized in silico analysis techniques. Here, we describe the selection of aptamers that can bind to AFP as well as in vitro and in silico validation of the binding site. We also developed an aptamer-based detection platform called hybrid-aptablotting sandwich assay (Hybrid-ABSA) and a point-of-care AFP detection kit.
Results
To construct the AFP detection platform, we screened a total of 18 candidate AFP-binding aptamers and identified a pair of aptamers without non-overlapping-binding sites using dot blotting. We validated results of dot blotting using in silico 3D binding structure docking simulation analysis. The aptamer-based Hybris-ABSA for AFP detection using this pair of aptamers demonstrated superior detection performance to antibody-based ELISA detection methods. Furthermore, we confirmed that AFP could be detected using the developed point-of-care lateral flow In Vitro Diagnostic (IVD) kit.
Conclusion
Results of this study demonstrate that aptamer-based disease diagnostic technology has potential of replace conventional diagnostic methods. This study also confirms that in silico techniques have sufficient utility in biological assays.
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Data availability
The data that support the findings of this study are available from the corresponding author upon reasonable request.
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Acknowledgements
This research was supported by Chungbuk National University Korea National University Development Project (2021).
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W-RS and D-YP: conceived and designed the study; H-JU, GA, and W-RS: performed data analysis; D-YP: performed the biological methodology and acquisition of data; SYK, J-YA, and Y-HK: drafted the manuscript (assign co-first authors order according to workload). All the authors read and approved the final version of the manuscript.
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Woo-Ri Shin declares that she has no conflict of interest. Dae-Young Park declares that he has no conflict of interest. Hyun-Ju Um declares that she has no conflict of interest. Gna Ahn declares that she has no conflict of interest. Sang Yong Kim declares that he has no conflict of interest. Ji-Young Ahn declares that she has no conflict of interest. Yang-Hoon Kim declares that he has no conflict of interest.
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Clinical samples were collected from the Institute of Clinical Medicine, Department of Internal Medicine, Chonbuk National University Hospital, South Korea. All subjects provided their written informed consent prior to participating in this study. The Ethics Committee and IRB (Institutional Review Board) of Chonbuk National University Hospital approved all experimental procedures.
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Shin, WR., Park, DY., Um, HJ. et al. 3D structural analysis of aptamer and diagnostic platforms for detecting hepatocellular carcinoma. Mol. Cell. Toxicol. 19, 621–634 (2023). https://doi.org/10.1007/s13273-023-00369-8
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DOI: https://doi.org/10.1007/s13273-023-00369-8