Detection of prostate specific antigen in whole blood by microfluidic chip integrated with dielectrophoretic separation and electrochemical sensing

The efficient detection of cancer markers has faced many challenges, such as severe interference, complicated and time-consuming operation, low sensitivity and so on. In this paper, a microfluidic chip integrated with electrodes for dielectrophoretic (DEP) separation, microchannels for electrochemical nanoprobes binding and differential pulse voltammetry (DPV) detection was proposed for the sensitive and rapid detection of prostate specific antigen (PSA) in whole blood. The functional units, which could realize cell separation, PSA derivatization (binding of electrochemical nanoprobes), capture and detection, were integrated on the microfluidic chip. The well-designed V-shaped interdigital electrode arrays provided DEP separation for blood cells with efficiency as high as 98%. Particularly, DEP effect significantly improved the sensitivity of PSA detection and reduced the detection limit by two orders of magnitude. In order to achieve sensitive detection of PSA, binding of electrochemical nanoprobes and then DPV detection was selected and integrated following the DEP separation. A sandwich structure based on electrochemical nanoprobes and dual-aptamers for on-chip DPV detection was proposed, which included self-synthesized electrochemical nanoprobes bovine serum albumin/detection aptamer 2/polythionine@gold nanoparticles (BSA/Apt2/PThi@Au NPs), target PSA, and sensing interface 6-mercaptohexanol/capture aptamer 1/gold nanoparticles on gold electrode (MCH/Apt1/Au NPs/Au). The method of quantitative detection of PSA in whole blood was then established. The excellent performance of the microfluidic chip allowed the determination of PSA in whole blood in the range of 1 pg/mL ∼10 ng/mL with an ultralow limit of detection of 0.25 pg/mL, which was better than the results obtained by conventional methods.