Graphene oxide as a cartridge enable on-line assembly of photosensitizer for .sup.1O.sub.2-based electrochemical aptasensing

An ultrasensitive .sup.1O.sub.2-based electrochemical aptasensor by on-line assembly of photosensitizers using graphene oxide (GO) as a cartridge is reported. In the presence of target protein lysozyme, the interaction of lysozyme with aptamer led to the dissociation of dsDNA and release of the aptamer-lysozyme complex to solution, with DNA-c retaining on the electrode; then, the photosensitizer phloxine B (PB) was assembled on the electrode since GO can simultaneously adsorb DNA-c and PB molecules. Upon irradiation by a green LED, .sup.1O.sub.2 was generated by photocatalysis of PB molecules and then cleaved the DNA-c, leading to remarkably decreased impedance signals that linearly respond with the logarithm of lysozyme concentration. Benefitting from the efficient photosensitization ability of PB and the high PB-loading capacity of GO, the developed sensor allowed determination of 0.001 to 100 nM lysozyme with a limit of detection of about 0.14 pM. The relative standard deviation (RSD) for five independent electrodes with 1 nM lysozyme was 3.1%, indicating satisfactory reproducibility. The sensor also showed excellent selectivity toward lysozyme in the presence of interfering substances and was applied to the determination of lysozyme in urine samples with recoveries ranging from 91 to 101%. The on-line assembly of photosensitizer technique opens a new way for amplified electrosensing of biomolecules.