Electrochemical Immunosensor for Cardiac Troponin I Detection Based on Covalent Organic Framework and Enzyme-Catalyzed Signal Amplification

Herein, a highly sensitive electrochemical immunosensor was presented for the cardiac troponin I (cTnI) determination using a multifunctional covalent organic framework-based nanocomposite (HRP-Ab2-Au-COF) as the signal amplification probe. The spherical COF with a large surface area was synthesized in a short time by a simple solution-based method at room temperature. The good biocompatibility, low toxicity, and high stability in water of the COF guarantee its application in biosensing. Besides, its high porosity makes it an excellent carrier for loading abundant horseradish peroxidase (HRP). The modified gold nanoparticles on the surface of COF not only provide a load platform for secondary antibody (Ab2) but also improve the conductivity of COF. Under the synergistic effect of the hydrogen peroxide (H₂O₂) and HRP, hydroquinone (HQ) in the solution is catalytically oxidized to benzoquinone (BQ), which is then reduced on the electrode surface to generate the electrochemical signal. The designed probes not only show the specific recognition behavior of Ab2 to cTnI but also improve the sensitivity of the biosensing system due to the signal amplification caused by the excellent enzyme catalytic performance of HRP. Based on the H₂O₂-HRP-HQ signal amplification system, the biosensor for cTnI was fabricated and exhibited a linear response as a function of logarithmic cTnI concentration ranging from 5 pg/mL to 10 ng/mL, and the detection limit was 1.7 pg/mL. Moreover, the biosensor exhibited excellent recovery and reproducibility in the actual sample testing. This work provided a simple approach to determine cTnI quantitatively in practical samples and broadened the utilization scope of the COF-based nanocomposite in the electrochemical immunosensor.