creators_name: Bharti, Bala creators_name: Bhardwaj, Priya creators_name: Bisht, Bhawana creators_name: Bala, Rajni creators_name: Sagrika, S creators_name: Rohit, Manojkumar creators_name: Singh, Jagtar creators_name: Bhalla, Vijayender type: article datestamp: 2026-07-13 08:15:14 lastmod: 2026-07-13 08:15:14 metadata_visibility: show title: Target-Induced Shielding Layer at the Dual-Affinity Probe Interface for Rapid Colorimetric Detection of Cardiac Biomarkers ispublished: pub subjects: QR keywords: nanoparticle shielding layer; rapid detection; colorimetric assay; cardiac troponin I (cTnI); microscale thermophoresis (MST) note: Copyright to this Article belongs to AMER CHEMICAL SOC abstract: Cardiac troponin I (cTnI) is a key biomarker for acute myocardial infarction (AMI); however, the available detection methods are limited by slow responses, complex steps, and costly instrumentation, restricting their suitability for rapid or point-of-care testing. Herein, we report a rapid and sensitive colorimetric assay for the detection of cTnI based on a nanoparticle shielding layer and its interfacial interactions with an enzyme. To achieve high sensitivity, we employed a dual-affinity silver nanoparticle (AgNP) probe coated with two highly specific affinity molecules, i.e., aptamer (Apt) and antibody (Ab), offering effective detection of the target molecule. The affinity layer coating was inherently imperfect, leaving exposed regions at the interface of the AgNP probe to inhibit the enzyme (urease) by Ag-SH interaction. Upon introduction of cTnI, the probe selectively binds to it, and a target-induced shielding layer is formed that prevents the interaction of the enzyme with the AgNP surface, rendering the enzyme active in solution. Consequently, the active enzyme generates a colored endpoint for visual detection in <15 min with a detection limit of 0.088 ng/mL. We assessed the assay performance in a complex matrix to evaluate the stringency. The working of the assay was studied using microscale thermophoresis (MST), which showed complete masking of the urease-particle interaction in the presence of cTnI. Unlike surface-based methods, such as ELISA, our solution-phase assay eliminates immobilization and blocking steps and achieves enhanced sensitivity, representing a paradigm shift in biomarker detection. date: 2026-03-18 date_type: published publication: ACS APPLIED MATERIALS & INTERFACES volume: 18 number: 8 publisher: AMER CHEMICAL SOC place_of_pub: 1155 16TH ST, NW, WASHINGTON, DC 20036 USA pagerange: 12423-12431 refereed: TRUE official_url: http://doi.org/10.1021/acsami.5c21395 citation: Bharti, Bala and Bhardwaj, Priya and Bisht, Bhawana and Bala, Rajni and Sagrika, S and Rohit, Manojkumar and Singh, Jagtar and Bhalla, Vijayender (2026) Target-Induced Shielding Layer at the Dual-Affinity Probe Interface for Rapid Colorimetric Detection of Cardiac Biomarkers. ACS APPLIED MATERIALS & INTERFACES, 18 (8). pp. 12423-12431. document_url: http://crdd.osdd.net/open/3470/1/references%20%283%29.bib