▎ 摘　　要

NOVELTY - Detecting GPC3 based on RGO-CS-Hemin/AuNPs nanocomposite materials involves forming RGO-CS-Hemin material, pouring graphene oxide into distilled water, using ultrasonic cell disruptor to sonic, making fully dissolved and uniform, and making GO aqueous solution, taking prepared GO aqueous solution and placing in beaker. The ascorbic acid is added to reduce GO to obtain RGO. The hemin is dissolved in ammonia water and diluted with distilled water to obtain hemin solution, mixed RGO with hemin. The hydrazine hydrate is added to the mixture of RGO and hemin for reduction, after water bath reaction, centrifuged to obtain RGO-Hemin complex. The reduced graphene oxide-chitosan-heme composite material is prepared by dissolving chitosan with acetic acid to obtain CS solution. USE - Method for detecting GPC3 based on RGO-CS-Hemin/AuNPs nanocomposite materials. ADVANTAGE - The method enables to detect GPC3 based on RGO-CS-Hemin/AuNPs nanocomposite materials that has simple detection method, low-cost and detection limit and saves time. DETAILED DESCRIPTION - Detecting GPC3 based on RGO-CS-Hemin/AuNPs nanocomposite materials involves forming RGO-CS-Hemin material, pouring graphene oxide into distilled water, using ultrasonic cell disruptor to sonic, making fully dissolved and uniform, and making GO aqueous solution, taking prepared GO aqueous solution and placing in beaker. The ascorbic acid is added to reduce GO to obtain RGO. The hemin is dissolved in ammonia water and diluted with distilled water to obtain hemin solution, mixed RGO with hemin. The hydrazine hydrate is added to the mixture of RGO and hemin for reduction, after water bath reaction, centrifuged to obtain RGO-Hemin complex. The reduced graphene oxide-chitosan-heme composite material is prepared by dissolving chitosan with acetic acid to obtain CS solution. The CS solution is added to RGO-Hemin complex, activated with EDC/NHS, and centrifuged to obtain RGO-CS-Hemin suspension. The modification of the electrode and construction of the biosensing interface screen-printed electrode are placed in the sulfuric acid solution and performs cyclic voltammetric scanning to obtain activated screen-printed electrode which is rinsed with water. The activated screen-printed electrode is placed in the chloroauric acid solution for constant potential deposition, after deposition is completed. The electrode is rinsed with water to obtain AuNPs/SPE electrode. The AuNPs/SPE electrode is soaked in glutaraldehyde, washed with PBS, dried, and then RGO-CS-hemin suspension is added dropwise to incubate, washed with PBS, and dried to obtain RGO-CS-Hemin/AuNPs/SPE electrode. The aminated GPC3 aptamer is taken and dropped on the sensor interface, incubated for period of time. The unfixed GPC3 aptamer is washed with PBS solution, and bovine serum protein solution is added dropwise to block to obtain GPC3aptamer/RGO-CS-Hemin/AuNPs/SPE sensor interface, dried for later use. The standard curve of GPC3 is drawn. The standard GPC3 solution is added to the GPC3aptamer/RGO-CS-Hemin/AuNPs/SPCE sensing interface obtained, incubated for period of time, washed with PBS solution to obtain working electrode, and dried for later use. The working electrode is placed in the PBS solution, DPV scan of the electrochemical workstation is used to record the peak current, respectively detects different concentrations of GPC3, drawn standard curve, and calculates the minimum detection limit of the method. The actual serum sample to be tested is washed, incubated for period of time, washed with PBS solution to obtain the working electrode, dried for later use. The working electrode is placed into the PBS solution, DPV scan of the electrochemical workstation is used to record the peak current. The concentration of GPC3 is calculated in the actual serum sample to be tested.