Fabrication of Polymeric Ferrocene Nanoparticles for Electrochemical Aptasensing of Protein with Target-Catalyzed Hairpin Assembly

Herein we describe a novel isothermal and enzyme-free electrochemical aptasensor for protein detection via the employment of polymeric ferrocene nanoparticles (PFcNPs) and target-catalyzed hairpin assembly amplification. The synthesized PFcNPs not only load numerous Fc molecules for enhanced electrochemical output but also possess plenty of amino groups, which increase the water solubility and facilitate the conjugation with the aptamer toward the recognition of target protein. After the formation of an aptamer/protein complex, the conformation of the DNA probe changes, which further triggers hairpin assembly on top of DNA tetrahedral structures modified on the electrode interface. The process can be recycled, and multiple PFcNPs are localized on the electrode. Thus, an amplified electrochemical signal is able to be recorded, which is sufficient to achieve a demonstrated limit of detection as low as 67 fM. This developed aptasensor can also discriminate target protein from other interfering substances with a high selectivity. Furthermore, it has been successfully applied in diluted real blood serum samples. All of these features make the present methodology a promising candidate for ultratrace protein biosensors.