Enhanced performance of an electrochemical aptasensor for real-time detection of vascular endothelial growth factor (VEGF) by nanofabrication and ratiometric measurement.

Achieving a biosensing interface without baseline drift caused by variables in matrix samples is essential for real-time detection of analytes. In this study, we developed a molecular beacon based electrochemical aptasensor to realize the ratiometric signal quantification of VEGF in serum by surface modification of nanocomposites of graphene oxide/methylene blue (GO/MB) and AuNPs followed by the attachment of ferrocene-labeled aptamer (aptamer-Fc) against VEGF. The presence of VEGF can trigger the configuration change of aptamer-Fc, resulting in the redox probe Fc being far away from the electrode surface to attenuate the electrochemical communication between electrode and Fc. Meanwhile, signal of MB also decreased due to the impediment of aptamer-Fc to electron transfer passage. The achieved GC-rGO/MB-AuNPs-aptamer-Fc sensing interface was successfully used for the sensitive detection of VEGF in real-time with a linear detection range 2-500 pg mL and detection limit of 0.1 pg mL based on ratiometric dual signal (Fc and MB) read-out. It was observed loading MB and AuNPs to the GO based sensing interface was favorable to enhance the analytical performance in terms of sensitivity and capability to effectively eliminate background interference. This electrochemical aptasensor provides a universal and reliable biosensing platform which is potential for real-time and sensitive tracking of various cytokines in vivo.