A facile electrochemical aptasensor for lysozyme detection based on target-induced turn-off of photosensitization

The quantification of proteins is essential in fundamental research or clinical applications. Here, we developed a facile electrochemical aptasensor based on target-induced turn-off of photosensitization for label-free and ultrasensitive detection of protein (exemplified by lysozyme). EB (ethidium bromide) molecules that were embedded in dsDNA between lysozyme binding aptamer and complementary DNA immobilized on the electrode, could photo-cleave the dsDNA via singlet oxygen (O₂¹) during photosensitization, resulting in a high voltammetry current of the [Fe(CN)₆]^3-/4-. Upon recognition of the lysozyme by aptamer, the EB molecules were released from dsDNA, and its photosensitization activity was turned off. As a result, more amount of complementary DNA was retained on the Au nanoparticles modified carbon nanotube paste electrode (AuNPs-CNPE), leading to a declined voltammetry current. Such a sensing strategy allowed detection of 10 pM-1 µM lysozyme with a low detection limit (about 2 pM). Besides, the sensor was free of labeling procedure as well as extra signal amplification step, and the CNPE modification was quite simple, only with AuNPs. The sensor also showed excellent selectivity toward lysozyme in the presence of interfering proteins, such as thrombin, bovine serum albumin, myoglobin, etc. The proposed sensor was applied to the determination of lysozyme in urine samples with the recoveries ranging from 96.6% to 101%. The proposed biosensor holds a great promise in developing other electrochemical sensors based on photosensitization.