A label-free aptasensor for highly sensitive detection of homocysteine based on gold nanoparticles

In the present study, an original electrode fabrication approach was devised to create a label free sensitive electrochemical aptasensor for the detection of Homocysteine (Hcy) (Homocysteine signal was used for detection). To bind certain targets, synthetic oligonucleotides used as aptamers (APs) were specifically selected. Aptamers are substitutes for antibodies for analytical devices because of their sensitivity and high affinity. In this study, Hcy-Binding-Aptamer (HBA) was grafted onto the surface of Au nanoparticles/Glassy Carbon Electrode (Au/GCE) in order to create an aptasensor. The effects of buffer concentration, buffer type, interaction time, and aptamer concentration were investigated and optimized. In addition, Differential Pulse Voltammetry (DPV) was implemented to identify homocysteine. Favorable performance was achieved at a detection limit of 0.01 μM (S/N = 3) and linear range 0.05-20.0 μM. Furthermore, the fabricated aptasensor displayed desirable stability and reproducibility. The developed electrochemical aptasensor was found to have reasonable selectivity for the detection of homocysteine in the presence of cysteine and methionine. Analysis of real samples showed good ability of the proposed homocysteine biosensor to provide sensitive, quick, easy, and cost effective measurement of homocysteine in human blood serum and urine samples.