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Ferrocene can be used as a peroxidase mimic to catalyze 3,3′,5,5′-tetramethylbenzidine or aniline oxidation in the presence of H2O2.

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Thrombin has a strong correlation to a variety of diseases, and it is of great significance to improve the sensitivity of thrombin detection. Herein, we report a biosensor for highly sensitive thrombin detection (17 fM) by combination of click chemistry and surface-initiated atom transfer radical polymerization (SI-ATRP). A sandwich structure is formed among thiol-modified aptamer-1, thrombin and azido-labeled aptamer-2. Then the initiators of SI-ATRP (propargyl α-bromoisobutyrate, PBiB) can be introduced to electrode surface via electrochemically mediated Cu(I)-catalyzed azide-alkyne cycloaddition (eCuAAC). Subsequently, SI-ATRP is activated based on the reaction that ascorbic acid reduced Mn(III)meso-tetra(4-carboxyphenyl)porphine (Mn(III)-TCPP) to Mn(II)mesotetra(4-carboxyphenyl)porphine (Mn(II)-TCPP), and numerous monomers are grafted to electrode surface via SI-ATRP. The limit of detection is enhanced about 4.9 × 10 3 -fold compared with previous strategy, the sensitivity of this biosensor can be improved and this method also have desirable stability and reproducibility.