▎ 摘　　要

NOVELTY - Method for dual-signal detection of golgi protein 73 (GP73) by sandwich electrochemical sensor based on hemachrome-reduced graphene oxide-trimanganese tetraoxide nano-enzyme for non-diagnostic purposes, involves detecting different concentrations of GP73 respectively, recording the peak current, drawing working curve according to relationship between the peak current and GP73 concentration and calculating minimum detection limit of the method, and detecting GP73 in actual samples by mixing normal human serum samples with GP73 standard solution to make a mixed solution and dripping to sensing interface, dripping detection probe, incubating and washing to obtain a working electrode and drying, putting the working electrode into phosphate-buffered saline (PBS) buffer, scanning with square-wave voltammetry (SWV), recording the response current value of the sensor and calculating the concentration of GP73 in the actual serum sample to be tested according to the working curve. USE - Method for dual-signal detection of GP73 by sandwich electrochemical sensor based on H-rGO-Mn3O4 nano-enzyme for non-diagnostic purposes. ADVANTAGE - The method is capable of generating two electrochemical signals with good linear relationship in the range of 0.01100 ng/mL, enabling dual-signal detection of GP73 with detection limit of 0.01 ng/mL and detection sensitivity of 2.441 microA/microM/cm2. DETAILED DESCRIPTION - Method for dual-signal detection of GP73 by sandwich electrochemical sensor based on hemachrome-reduced graphene oxide-trimanganese tetraoxide (H-rGO-Mn3O4) nano-enzyme for non-diagnostic purposes, involves preparing hemachrome-reduced graphene oxide by mixing prepared reduced graphene oxide and hemin, in the ratio of 1:1, reacting in water bath and washing by centrifugation, preparing hemachrome-reduced graphene oxide-trimanganese tetraoxide-aptamer 1 (H-rGO-Mn3O4-Apt1) detection probe by adding manganese chloride tetrahydrate powder and polyvinylpyrrolidone to ultrapure water, stirring, heating, adding sodium hydroxide solution, stirring, centrifuging and washing to obtain trimanganese tetraoxide solution, adding N-hydroxysuccinimide (NHS)/1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) to the hemachrome-reduced graphene oxide solution, adding trimanganese tetraoxide under stirring conditions at room temperature, stirring, centrifuging and washing to obtain H-rGO-Mn3O4 nano-enzyme, taking Apt1 and H-rGO-Mn3O4 nanoenzyme solution and mixing with ultrasonic vibration, incubating, centrifuging and washing to obtain H-rGO-Mn3O4-Apt1 detection probe, performing construction of electrochemical sensing interface by placing screen printing electrode (SPE) in sulfuric acid solution, scanning by cyclic voltammetry (CV), washing and drying to obtain activated SPE, mixing o-phenylenediamine (OPD) with chloroauric acid, sinking the activated SPE into the mixed solution, using i-t technology for constant potential deposition, washing and drying to obtain nano-gold particle-doped poly-o-phenylenediamine/SPE (Au@POPD/SPE), dripping EDC/NHS was on the surface of Au@POPD/SPE for activation, dripping aptamer 2 (Apt2) to Au@POPD/SPE, incubating, washing and blow drying to obtain Apt2/Au@POPD/SPE, adding bovine serum albumin (BSA) solution to Apt2/Au@POPD/SPE surface, incubating, blocking non-specific binding sites, washing and drying, drawing GP73 standard curve by dripping standard GP73 sample on the electrochemical sensing interface and incubating to obtain GP73/Apt2/Au@POPD/SPE, dripping H-rGO-Mn3O4-Apt1 solution on the surface of GP73/Apt2/Au@POPD/SPE and incubating to obtain the working electrode H-rGO-Mn3O4-Apt1/GP73/Apt2/Au@POPD/SPE, putting the prepared working electrode into PBS buffer containing hydrogen peroxide and 1,2,4-trimethylbenzene, scaning with SWV and recording the response current change of the sensor, detecting different concentrations of GP73 respectively, recording the peak current, drawing working curve according to relationship between the peak current and GP73 concentration and calculating minimum detection limit of the method, and detecting GP73 in actual samples by mixing normal human serum samples with GP73 standard solution to make a mixed solution and dripping to the sensing interface, dripping detection probe, incubating and washing to obtain a working electrode and drying, putting the working electrode into the PBS buffer, scanning with SWV, recording the response current value of the sensor and calculating the concentration of GP73 in the actual serum sample to be tested according to the working curve. DESCRIPTION OF DRAWING(S) - The drawing shows a schematic view explaining the method for dual-signal detection of GP73 by sandwich electrochemical sensor based on H-rGO-Mn3O4 nano-enzyme for non-diagnostic purposes. (Drawing includes non-English language text).