Ultrasensitive simultaneous detection of four biomarkers based on hybridization chain reaction and biotin-streptavidin signal amplification strategy

A sandwich-type electrochemical immunosensor based on redox probe tags identification technology for ultrasensitive simultaneous detection of four antigens was proposed. In this project, well-distributed graphene/gold (GR-Au) hybrid film was acquired through one-step codeposition in an electrode surface and served as the base substrate for immobilizing capture antibodies (Ab1). Hybridization chain reaction (HCR) and biotin/streptavidin (B/SA), combining with gold magnetic nanoparticles were applied to increase the immobilization amount of signal tags in detection antibody (Ab2) bioconjugates. To verify this strategy, four representative biomarkers, a-fetoprotein (AFP), carcinoembryonic antigen (CEA), carbohydrate antigen (CA)125 and prostate special antigen (PSA), were used as model analytes. The resulting immunosensor could simultaneously detect four antigens in single-pass differential pulse voltammetry (DPV) scan, and exhibited obviously improved senstivity compared to previous similar immunosensors, displayed good linear relationships in the ranges from 0.2 to 800 pg/mL for AFP, 0.2 to 600 pg/mL for CEA, 0.2 to 1000 pg/mL for CA125, 0.2 to 800 pg/mL for PSA and with detection limits of 62, 48, 77 and 60 fg/mL, respectively.