Thread-based colorimetric biosensor for urea determination in serum Lewinska, Izabela Bącal, Paweł Orbe Payá, Ignacio De Capitán Vallvey, Luis Fermín Erenas Rodríguez, Miguel María Urea Microfluidic thread-based analytical device Nanoflowers Ionophore-based optical sensor Urea is a crucial biomarker in clinical analysis, mainly used to diagnose and monitor kidney condition. However, its reliable determination in serum in point-of-care testing format still remains a vital challenge. Here, we tackled this issue by developing a microfluidic thread-based optical biosensor for serum urea determination. The working principle of the presented device was combining enzymatic urea hydrolysis using urease with the resulting ammonium ions detection using ionophore-chromoionophore chemistry. Urease was immobilized on thread with the aid of synthesized urease-calcium phosphate nanoflowers while components for ammonium ions detection were embedded in PVC-based membrane located on the same thread. In the first step, ammonium ions determination in a thread-based sensor was optimized. Then, urease-calcium posphate nanoflowers (U-CaNFs) were included on the thread to ensure selectivity for urea. U-CaNFs synthesis was optimized and the resulting nanoflowers were characterized using various analytical techniqes (e.g. SEM, EDS, TGA, XRD). The calculated limit of detection for urea was 37 µmol L−1 and the total analysis time was only 8 min. The developed thread-based devices were validated with control sera samples, proving their high accuracy. 2026-02-24T07:48:16Z 2026-02-24T07:48:16Z 2026-05-15 journal article Sensors and Actuators: B. Chemical 455 (2026) 139644 https://hdl.handle.net/10481/111408 https://doi.org/10.1016/j.snb.2026.139644 eng http://creativecommons.org/licenses/by-nc-nd/4.0/ open access Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License Attribution-NonCommercial-NoDerivatives 4.0 Internacional