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Biosens. Bioelectron., Available online 14 May 2025 | https://doi.org/10.1016/j.bios.2025.117584
Gold nanoparticle-facilitated assembly via supernatant transfer as a colorimetric SELEX and sensing platform for cortisol
Duc Loc Sai a,1, Jeong Min Lee a,1, Kyung-Min Kim a, Yeonju Lee a, Seoyeon Choi b,c, Chanmi Kim b,c, Hyo-Il Jung b,c,d, Young-Pil Kim a,e,f,g
aDepartment of Life Science, Hanyang University, Seoul 04763, Republic of Korea
bSchool of Mechanical Engineering, Yonsei University, Seoul 03722, Republic of Korea
cThe DABOM Inc., Seoul 03722, Republic of Korea
dDepartment of Integrated Medicine, Yonsei University, Seoul 03722, Republic of Korea
eResearch Institute for Convergence of Basic Science, Hanyang University, Seoul 04763, Republic of Korea
fResearch Institute for Natural Sciences, Hanyang University, Seoul 04763, Republic of Korea
gHanyang Institute of Bioscience and Biotechnology, Hanyang University, Seoul 04763, Republic of Korea
1These authors contributed equally to this work.
Corresponding authors: Hyo-Il Jung, Young-Pil Kim
aDepartment of Life Science, Hanyang University, Seoul 04763, Republic of Korea
bSchool of Mechanical Engineering, Yonsei University, Seoul 03722, Republic of Korea
cThe DABOM Inc., Seoul 03722, Republic of Korea
dDepartment of Integrated Medicine, Yonsei University, Seoul 03722, Republic of Korea
eResearch Institute for Convergence of Basic Science, Hanyang University, Seoul 04763, Republic of Korea
fResearch Institute for Natural Sciences, Hanyang University, Seoul 04763, Republic of Korea
gHanyang Institute of Bioscience and Biotechnology, Hanyang University, Seoul 04763, Republic of Korea
1These authors contributed equally to this work.
Corresponding authors: Hyo-Il Jung, Young-Pil Kim
Abstract
Accurate and selective detection of cortisol, a key biomarker for stress and metabolic health, is essential for diagnostics and healthcare. Single-stranded DNA (ssDNA) aptamers hold great promise for small molecule detection, yet conventional systematic evolution of ligands by exponential enrichment (SELEX) methods are often hindered by invisible screening and restricted conjugation capabilities. Here, we report a rapid and straightforward approach for cortisol aptamer discovery and determination using gold nanoparticle-facilitated assembly via supernatant transfer (GNP-FAST) coupled with GNP-SELEX. These GNP-mediated visual processes enable efficient ssDNA library refinement in only 6 rounds of GNP-FAST, followed by rapid aptamer discovery within 12 rounds of GNP-SELEX, culminating in a highly selective colorimetric assay for cortisol. Comprehensive engineering and characterization, including molecular docking, circular dichroism spectroscopy, and isothermal titration calorimetry, validated the performance of the identified aptamers. Notably, a truncated 26-mer of the selected aptamer, CorApt5-T, exhibited high binding affinity (KDapp≈210 nM) and specificity for cortisol through positive cooperativity on the GNP surface. Consequently, CorApt5-T enabled a simple and rapid GNP-based colorimetric assay for cortisol in saliva and serum, demonstrating more pronounced cortisol-driven structural changes and superior sensitivity when compared to previously reported cortisol aptamers. These findings demonstrate the potential of integrating GNP-based colorimetric assay with GNP-FAST and GNP-SELEX as a visible platform for developing biosensors targeting a broad range of small molecules.
논문정보
- Research article
- 2025년 05월 (BRIC 등록일 2025-06-11)
- 국내 연구진
- 바이오·의료융합 > 바이오센싱 및 나노바이오물질
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