Development of a one-shot dual aptamer-based fluorescence nanosensor for rapid, sensitive, and label-free detection of periostin

www.nature.com/scientificreports OPEN Development of a one‑shot dual aptamer‑based fluorescence nanosensor for rapid, sensitive, and label‑free detection of periostin Jonghoon Park & Changill Ban * Periostin is associated with several diseases, including cancers. Therefore, monitoring blood periostin levels is a powerful tool for diagnosing various diseases and identifying their severity. However, conventional detection methods pose several challenges, including high costs. To address these issues, we developed a novel one-shot dual aptamer-based fluorescence nanosensor for detecting periostin. The proposed nanosensor facilitates rapid, label-free, and sensitive detection of periostin using gold nanoprobes constructed by rhodamine-b isothiocyanate, PL2trunc aptamer, and gold nanoparticles and silver nanoprobes fabricated by the PL5trunc aptamer and silver nanoparticles. The two nanoprobes form a core-satellite structure by interacting with periostin, and the nanosensor detects periostin through the fluorescence regenerated by the increased proximity between them. The nanosensor successfully detected periostin with remarkable detection limits of 106.68 pM in buffer and 463.3 pM in serum-spiked conditions within 30 min without additional washing or signal amplification processes. Considering serum periostin levels in various diseases, the proposed nanosensor provides a suitable method for identifying patients with various diseases and determining disease severity. Moreover, the platform can be helpful as a practical method for on-site medical diagnosis because it can be adapted to detect other biomarkers simply by replacing the aptamer with other detection probes. Monitoring blood biomarkers is a powerful tool for the diagnosis and prognosis of diseases such as cancer and diabetes1. Blood is a unique pathological sample as it can be obtained without causing much pain to the patient and carries several forms of disease biomarkers, such as proteins, nucleic acids, tumor cells, and m etabolites2,3. Among them, protein biomarkers are considered the most useful, as they are directly connected to cellular functions and can be easily t argeted3,4. Periostin is an extracellular matrix protein involved in various pathophysiological processes, including cell proliferation and cancer p athogenesis5,6. The periostin level in the blood is elevated in patients with diseases such as non-small cell lung cancer, obstructive sleep apnea-hypopnea syndrome, and diabetic retinopathy7–9. Therefore, highly sensitive methods for detecting periostin in the blood are in demand for the diagnosis and prognosis of these diseases. Conventional and widely used detection methods for protein biomarker include mass spectroscopy, surfaceenhanced Raman scattering (SERS), microarray, and enzyme-linked immunosorbent assay (ELISA)10–14. These methods are immunoassay-based utilizing antibodies and enzymes and have high sensitivity and selectivity4. However, they have several limitations, including the need for expensive equipment and experts, complicated steps such as signal amplification and washing, and high costs15. Moreover, cost-effective and sensitive alternative detection systems for periostin have not yet been developed. Consequently, the development of new molecular probes and detection systems that can replace antibodies for economical, simple, and rapid detection is desired. Nanotechnology presents an innovative way to overcome the limitations of conventional methods using small materials, including aptamers and metal nanoparticles. Aptamers are single-stranded oligonucleotides (ssDNA) that can specifically bind to a target molecule owing to their three-dimensional structures16. Aptamers can be developed from a randomized pool of libraries using the systematic evolutions of ligands by exponential Department of Chemistry, Pohang University of Science and Technology, 77, Cheongam‑ro, Nam‑gu, Pohang, Gyeongbuk 37673, Republic of Korea. *email: Scientific Reports | (2023) 13:10224 | https://doi.org/10.1038/s41598-023-37418-0 1 Vol.:(0123456789) www.nature.com/scientificreports/ enrichment (SELEX)17. Aptamers offer many advantages over antibodies, including large-scale synthesis, low cost, no batch-to-batch variation, and high chemical and thermal s tability18,19. Owing to these advantages, aptamers are widely applied in developing on-site sensors, modularization, and convenient multifunctional b iosensors20. Meanwhile, metal nanoparticles (NPs) can act as quenchers (~ 5 nm distance) or enhancers (10–90 nm distance) depending on their distance from the fluorophore due to their Förster resonance energy transfer (FRET) and metal-enhanced fluorescence (MEF) properties by a localized surface plasmon resonance (SPR) e nergy21. In addition, they have a high surface-to-volume r atio22. Moreover, metal NPs can act as a fluorescence regenerator in which the MEF effect of one NP can regenerate the quenched fluorescence of the fluorophore conjugated to another NP when both NPs form a core-satellite s tructure23. Therefore, metal NPs have been widely applied in various fields, including bioimaging, biosensors, and biomedical a pplications24. In this study, we report the development of a rapid and label-free one-shot periostin detection platform with high sensitivity and selectivity using aptamers, rhodamine-b isothiocyanate (RiTC), and metal NPs (Fig. 1). By adding gold (Au) and silver (Ag) nanoprobes at once, the nanosensor provides rapid and direct detection of periostin without additional washing or signal amplification processes. For fabricating the nanosensor, we first developed two periostin-specific ssDNA aptamers ( PL2trunc and P L5trunc) and then applied them to fabricate the two nanoprobes. For synthesizing the Au nanoprobes, Au@RiTC NPs constructed by attaching RiTC to AuNPs and PL2trunc aptamer were conjugated by Au-thiol interaction. Similarly, the Ag nanoprobes were synthesized by attaching the PL5trunc aptamers to AgNPs by Ag-thiol interaction. The two aptamers allow the stable dispersion of both nanoprobes in solutions and provide high specificity for periostin. In addition, they play an essential role in increasing the proximity between the two nanoprobes because they have a relatively smaller size than antibodies and a simultaneous binding property to periostin. The fluorescence of RiTC on the AuNP surface is quenched by the FRET effect induced by the direct conjugation of RiTC and AuNPs, which decreases the noise of the platform. Moreover, in the presence of periostin, the two nanoprobes form core-satellite structures, which results in the fluorescence regeneration of the quenched RiTC by AuNPs through the MEF effect of AgNPs. Therefore, the two nanoprobes facilitate rapid one-shot detection of periostin without the need for any signal Figure 1.  Detection strategy of periostin using the dual aptamer-based fluorescence nanosensors. The nanosensor uses Au nanoprobes constructed by Ri (...truncated)