The platinum nanozymes capped with tyrosine (Pt-Tyr NZ) were synthesized which demonstrated peroxidase and oxidase catalytic activities toward 3,3',5,5'-tetramethylbenzidine (TMB). The kinetic analyses revealed that Pt-Tyr NZ exhibited higher catalytic activity and substrate affinity for both H2O2 and TMB compared to horseradish peroxidase (HRP), indicating its superior efficiency in biosensing applications. The synthesized Pt-Tyr NZ was reversibly conjugated with complementary DNA (cDNA), which can selectively hybridize with miRNA-21, a cancer biomarker, and after hybridization with miRNA-21, detaches from the surface of Pt-Tyr NZ. Upon cDNA conjugation, the catalytic activity of Pt-Tyr NZ decreased, but it was restored in the presence of miRNA-21, providing a one-step spectrophotometric biosensing method for miRNA-21 determination. Optimization of parameters affecting the sensitivity of the developed method for miRNA-21 detection was performed, including the tyrosine-to-platinum ratio in Pt-Tyr NZ synthesis, Pt-Tyr NZ concentration, and cDNA concentration for conjugation. The developed method was fully validated and showed promising results in terms of reproducibility (RSD = 1.26%, 95% confidence interval = 0.03, n = 6), repeatability (RSD = 0.83%, n = 5), and accuracy for spiked miRNA-21 determination in 10% filtered human serum samples (recovery rate = 109.7 ± 4.2%). It showed a linear dynamic range (LDR) from 37.1 to 185 nM, with a limit of detection (LOD) of 11.1 nM and a limit of quantification (LOQ) of 36.9 nM.

Bioinspired Materials and Biosensor Technologies Institute of Materials Science Faculty of Engineering Kiel University Kiel 24143 Germany

Central European Institute of Technology Brno University of Technology Brno 61200 Czech Republic

Department of Chemistry and Biochemistry Mendel University in Brno Brno 61300 Czech Republic

Institute of Photonics and Electronics Czech Academy of Sciences Prague 18200 Czech Republic

Kiel Nano Surface and Interface Science Kiel University institution Kiel 24118 Germany

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