CN111307908A - Method for detecting GPC3 based on H-rGO-Pt @ Pd NPs nano composite material - Google Patents
Method for detecting GPC3 based on H-rGO-Pt @ Pd NPs nano composite material Download PDFInfo
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Abstract
A method for detecting GPC3 based on H-rGO-Pt @ Pd NPs nano composite material. The method comprises the steps of preparing an H-rGO-Pt @ Pd NPs material, modifying an electrode, constructing a biosensing interface, drawing a standard curve of GPC3, and detecting an actual sample. By constructing a H-rGO-Pt @ Pd NPs/Au NPs/SPCE biosensing platform, the biosensor can be specifically combined with a GPC3 aptamer by using GPC3, and electrochemical signals in PBS (phosphate buffer solution) solutions before and after GPC3 are detected by a DPV (differential pressure voltammetry) method, so that the detection of GPC3 is realized. The method has the advantages of simple operation, time saving, low cost and lower detection limit.
Description
Technical Field
The invention belongs to the field of biological detection, and particularly relates to a method for specifically detecting GPC3 based on a nano composite material.
Background
Glypican-3 (GPC 3) is a novel protein which has been recently discovered and is most promising as a tumor marker specific for HCC. At present, enzyme-linked immunosorbent assay (ELISA), serum tumor markers, radioimmunoassay and the like are mainly adopted in a GPC3 detection method. The invention patent of publication No. CN 105717104B discloses a peripheral blood GPC3 detection method for hepatocellular carcinoma patients: separating and acquiring CTC in peripheral blood of a liver cancer patient, which cannot obtain a tissue specimen, by using a membrane filter device, manufacturing a thin layer slice by using a cell wax block technology, and further detecting the expression condition of GPC 3. The invention patent of publication No. CN 110872351A relates to a nano antibody GN1 specifically binding GPC3 protein, and a preparation method and application thereof. The invention patent with publication number CN 101290318B discloses an ELISA kit for diagnosing liver cancer. The instruments used in the methods are expensive, the operation is complex, the detection sensitivity is low, radioactive pollution exists, and a rapid, sensitive and easy-to-operate GPC3 detection method needs to be established.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for detecting GPC3 based on a nano composite material of heme/reduced graphene oxide/platinum @ palladium (H-rGO-Pt @ Pd NPs), so that the sensitivity is improved, and the specificity is enhanced.
In order to solve the technical problem, an electro-deposition technology and an electrostatic adsorption effect are adopted to manufacture the GPC3 nano aptamer electrochemical biosensor based on H-rGO-Pt @ Pd NPs/Au NPs. The peak current of the graphene and nano-metal is recorded by Differential Pulse Voltammetry (DPV) of an electrochemical workstation by utilizing the high loading capacity and good electron transfer effect of the graphene and nano-metal on a GPC3 aptamer and the specific recognition effect of a GPC3 aptamer on GPC 3. The incubation temperature of GPC3, the concentration of the composite, the GPC3 aptamer concentration, the incubation time, the amount of composite, and the pH of PBS were optimized and a standard curve was plotted, giving the correct GPC3 concentration by comparison to a standard working curve. Compared with the existing method, the method has the advantages of relatively simple operation, high specificity, less time and cost consumption, and capability of reaching the minimum detection limit of 0.1845 ng/mL.
The detection principle of the invention is as follows: H-rGO-Pt @ Pd NPs/AuNPs are modified on the surface of a screen printing electrode by adopting an electrodeposition technology and a glutaraldehyde crosslinking method. The AFP aptamer is loaded on the surface of an H-rGO-Pt @ Pd NPs/Au NPs material through a nanotechnology and intermolecular force, and the aptamer exists in a biosensing interface in a single-chain structure form due to an unstable spatial structure of the aptamer. After GPC3 is added on a biosensing interface, GPC3 can be specifically combined with GPC3 aptamer to generate a stable spatial structure, so that the spatial structure can be orderly arranged on the surface of an electrode. The detection of GPC3 was achieved by detecting the electrochemical signal (scan rate 0.05V/s, voltage interval-0.4V-1.2V) in PBS solution (0.2 mol/L, pH 6.0) before and after GPC3 by the DPV method and plotting the current versus the concentration of GPC 3.
The invention is carried out according to the following steps:
step 1: preparation of H-rGO-Pt @ Pd NPs material
(1) Preparation of Reduced Graphene Oxide (RGO): pouring Graphene Oxide (GO) into distilled water, and performing ultrasonic treatment by using an ultrasonic cell disruption instrument to fully and uniformly dissolve the Graphene Oxide (GO) to prepare a GO aqueous solution. And (3) putting the GO aqueous solution into a beaker, and adding Ascorbic Acid (AA) to reduce GO to obtain RGO.
(2) Preparation of heme/reduced graphene oxide (H-rGO): dissolving heme in ammonia water to obtain heme solution. And (3) mixing the heme solution with the Reduced Graphene Oxide (RGO) solution, mixing with a hydrazine hydrate solution, and stirring to obtain an H-rGO mixed solution.
(3) Preparation of a heme/reduced graphene oxide/platinum @ palladium (H-rGO-Pt @ Pd NPs) composite material: adding phthalic acid diethylene glycol diacrylate (PDDA) and sodium chloride (NaCl) into the H-rGO solution, and stirring for reaction to obtain the PDDA modified H-rGO solution. Mixing sodium chloroplatinate (Na)2PtCl6) And sodium tetrachloropalladate (Na)2PdCl4) Adding the mixture into a PDDA modified H-rGO solution, stirring for reaction, then adding an Ethylene Glycol (EG) solution into the solution for mixing, adjusting the pH value of the mixed solution by using sodium hydroxide (NaOH), and carrying out reflux reaction. And centrifugally cleaning to obtain the H-rGO-Pt @ Pd NPs composite nano material.
Step 2: electrode modification and biosensing interface construction
(1) Placing a Screen printing electrode (SPCE) at H2SO4In solution, intoAnd (5) performing cyclic voltammetry scanning to obtain an activated screen-printed electrode, and washing with water.
(2) And (3) placing the activated screen printing electrode into a chloroauric acid solution, carrying out constant potential deposition, and washing the electrode clean by water after the deposition is finished to obtain the Au NPs/SPCE electrode.
(3) And soaking the Au NPs/SPCE electrode with glutaraldehyde, washing with PBS, drying, then dropwise adding H-rGO-Pt @ PdNPs suspension, incubating for a period of time, washing with PBS, and drying in the air to obtain the H-rGO-Pt @ Pd NPs/Au NPs/SPCE electrode.
(4) Taking carboxylated GPC3 aptamer (GPC 3)Apt) Adding dropwise into sensor interface, incubating for a period of time, washing GPC3 aptamer which is not fixed on the interface with PBS solution, adding Bovine Serum Albumin (BSA) solution, and blocking to obtain GPC3AptAnd the/H-rGO-Pt @ Pd NPs/Au NPs/SPCE sensing interface is dried for later use.
And step 3: GPC3 Standard Curve
(1) Dropwise adding standard GPC3 solution to GPC3 obtained in step 2AptAnd incubating the/H-rGO-Pt @ Pd NPs/Au NPs/SPCE sensing interface for a period of time, washing the sensing interface with a PBS solution to obtain a working electrode, and airing the working electrode for later use.
(2) The working electrode was placed in PBS solution and its peak current was recorded using DPV scanning at the electrochemical workstation.
(3) GPC3 was detected at different concentrations, and a calibration curve was plotted to calculate the minimum detection limit of the method.
And 4, step 4: detection of actual samples
(1) GPC3 obtained in step 2AptAnd (2) dripping an actual sample to be detected on a/H-rGO-Pt @ Pd NPs/Au NPs/SPCE sensing interface, incubating for a period of time, cleaning with a PBS solution to obtain a working electrode, and airing for later use.
(2) The working electrode was placed in PBS solution and its peak current was recorded using DPV scanning at the electrochemical workstation.
(3) And (4) obtaining the concentration of GPC3 in the actual sample to be tested according to the standard curve in the step 3.
Further, the concentration of the GO solution in the step 1 is 1.0 mg/mL.
Further, the ammonia aqueous solution in the step 1 is 10 μ L80%.
Further, the hydrazine hydrate solution in the step 1 is 8 μ L.
Further, the concentration of the H-rGO solution in the step 1 is 1.0 mg/mL.
Further, the PDDA in step 1 was 2mL, 0.2%.
Further, the amount of NaCl in the step 1 is 5mL and 0.2 mol/L.
Further, Na in said step 12PtCl6The concentration was 2mL and 20 mmol/L.
Further, Na in said step 12PdCl4The concentration was 2mL and 20 mmol/L.
Further, NaOH in the step 1 is 1 mol/L.
Further, EG in step 1 was 10 mL.
Further, the prepared materials are mixed in the step 1, the pH value of the mixed solution is adjusted by NaOH, and reflux reaction is carried out. And centrifugally cleaning to obtain the H-rGO-Pt @ Pd NPs composite nano material.
Further, the concentration of the H-rGO-Pt @ Pd NPs solution in the step 1 is 0.5 mg/mL.
Further, H in the step 22SO4The concentration of the solution was 0.5 mol/L.
Further, the scanning voltage in the step 2 is-0.4V-1.2V, and the number of scanning segments is 20.
Further, the electrode is placed in H in the step 22SO4After cyclic voltammetry scanning, the electrode is washed clean by pure water, then is put into chloroauric acid solution to be respectively subjected to cyclic voltammetry scanning, and finally is washed by pure water and dried for standby.
Further, in the step 2, the concentration of chloroauric acid is 0.01%, the deposition condition is-0.5V, and the deposition time is 120 s.
Further, in the step 2, the concentration of glutaraldehyde is 2.5%.
Further, the BSA solution had a concentration of 0.5%, PBS at a concentration of 0.2mol/L, and pH at 7.0.
Further, the GPC3 aptamer concentration in step 2 was 5. mu. mol/L.
Further, the GPC3 aptamer was incubated at 25 ℃ for 1 hour at the electrode.
Further, in step 2, the concentration of BSA was 0.5%.
Further, the optimum incubation temperature of GPC3 in step 3 is 15 ℃, and the optimum incubation time is 60 min.
Preferably, the linear scanning range in step 3 and step 4 is-0.4V-1.2V, and the scanning rate is 0.05V/s.
Wherein the unique H-rGO-Pt @ Pd NPs nano composite material prepared in the step 1 is GPC3AptProvides an excellent carrier. Step 2 constitutes a biosensing interface that specifically recognizes GPC3 and facilitates the transfer of electrons. The construction of the biosensing interface in step 2 is an essential key step in the electrochemical detection of GPC3 in step 3. It can be seen that steps 1-3 are mutually supported and act together to realize the detection of GPC3 by using H-rGO-Pt @ Pd NPs composite material and GPC3 aptamer as recognition probes.
Compared with the prior art, the invention has the following advantages:
1. the H-rGO-Pt @ Pd NPs composite material has the advantages of large specific surface area, good biocompatibility, strong surface reaction activity, high catalytic efficiency and strong electron transfer capacity; the specific surface areas of the nano platinum, the nano palladium and the nano graphene are large, the biocompatibility is good, the electron transfer capacity is strong, the load capacity of fixed biomolecules can be increased, the electron transfer between an electrode and a biological component is promoted, a GPC3 aptamer can be effectively fixed on the surface of the electrode, the signal conduction is accelerated, the stability of a sensor is ensured, and the detection capacity is improved; GPC3 can perform specific binding reaction with GPC3 aptamer to generate stable spatial structure. Compared with the traditional sensor, the novel nano material sensor has the advantages of smaller volume, higher speed, higher precision and higher reliability.
2. The background interference of GPC3 detected by adopting a GPC3 aptamer as a recognition probe is small, and the detection limit of 0.1845ng/mL can be reached. The affinity between the aptamer and the target is often stronger than the affinity between the antigen and the antibody. Furthermore, aptamers are more easily labelled and modified by chemical means than antibodies, and these treatments facilitate the functionalization of nanoparticles and their surfaces.
Drawings
FIG. 1 is a schematic diagram of H-rGO-Pt @ Pd NPs based nano-aptamer sensor detection GPC 3;
FIG. 2 transmission electron micrograph of H-rGO-Pt @ Pd NPs composite nanomaterial;
FIG. 3 DPV curves for different GPC3 concentrations;
FIG. 4 is a working curve of GPC 3.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
A method for detecting GPC3 based on a nano composite material of heme/reduced graphene oxide/platinum @ palladium (H-rGO-Pt @ Pd NPs) is disclosed, and the detection principle is shown in figure 1. First, H2SO4And (4) activating the bare electrode, and performing electrodeposition on the gold nanoparticles on the surface of the bare electrode. Fixing the H-rGO-Pt @ Pd NPs nano composite material on the surface of Au NPs/SPCE by a glutaraldehyde crosslinking method to obtain an H-rGO-Pt @ Pd NPs/Au NPs/SPCE biosensor platform, loading a GPC3 aptamer on the surface of the H-rGO-Pt @ Pd NPs/Au NPs/SPCE material by a nanotechnology and intermolecular force, wherein the aptamer exists on the surface of the composite material in a single-chain structure due to an unstable spatial structure, and then blocking unreacted non-specific sites by bovine serum albumin. After GPC3 is added into a biosensing interface, GPC3 can be specifically combined with GPC3 aptamer to generate a stable spatial structure, so that the space can be orderly arranged on the surface of an electrode. The detection of GPC3 was achieved by detecting the electrochemical signal (scan rate 0.05V/s, voltage interval-0.4V-1.2V) in PBS solution (0.2 mol/L, pH 6.0) before and after GPC3 by the DPV method and plotting the current versus the concentration of GPC 3.
The specific implementation steps are as follows:
1. preparation of H-rGO-Pt @ Pd NPs composite nano material
(1) Weighing 30mg of graphene oxide, placing the graphene oxide in a beaker, adding 30 mL of ultrapure water, ultrasonically crushing the graphene oxide by using an ultrasonic cell crusher to fully and uniformly dissolve the graphene oxide to obtain a solution with the concentration of 1 mg/mL, taking 10mL of the uniformly dissolved solution in the beaker, adding 10 mg of ascorbic acid, placing the solution in a constant-temperature magnetic stirrer, and stirring at room temperature for 12 hours to obtain Reduced Graphene Oxide (RGO). FIG. 2A is a transmission electron micrograph of RGO showing a black lamellar structure, which shows the formation of a new reduced graphene oxide particle.
(2) Dissolving 30mg of heme in 10 mu L of ammonia water, adding 30 mL of ultrapure water into the solution, stirring the solution by using a glass rod to fully dissolve the heme to obtain a solution with the concentration of 1 mg/mL, mixing the heme solution and the reduced graphene oxide solution according to the proportion of 1:1, adding 8 mu L of hydrazine hydrate solution into the mixed solution, whirling the mixed solution for 10 min, putting the mixed solution into a water bath kettle at the temperature of 60 ℃ for water bath for 4H, then centrifuging the mixed solution in a centrifuge at the rotating speed of 13500 r/min for 10 min, removing supernatant, washing twice and drying to obtain the H-rGO composite nanomaterial. FIG. 2B is a transmission electron micrograph of H-rGO, and the black lamellar structure successfully adsorbs the particle particles, indicating that the H-rGO material is successfully constructed.
(3) Adding 2mL of PDDA (0.2%) and 5mL of NaCl (0.2M) into 10mL of H-rGO (0.5 mg/mL) solution, stirring for reaction for 12H, and carrying out centrifugal washing to obtain a PDDA modified H-rGO solution. 2mL of Na2PtCl6(20 mM) and 2mL Na2PdCl4(20 mM) was added to a PDDA modified H-rGO (0.5 mg/mL) solution and stirred for 12H, then 10mL of ethylene glycol solution was added to the solution and mixed, and the pH of the mixed solution was adjusted to 12 with 1M NaOH and refluxed at 140 ℃ for 4H. And centrifugally cleaning to obtain the H-rGO-Pt @ Pd NPs composite nano material. FIG. 2C is a transmission electron microscope image of H-rGO-Pt @ Pd NPs, and compared with FIG. 2B, more spherical small particles are randomly distributed on the surface of a black film, so that the successful construction of the H-rGO-Pt @ PdNPs material is proved.
2. Electrode modification and biosensing interface construction
(1) The screen-printed electrode (SPCE) was first soaked at 0.5 mol/L H before use2SO4Performing Cyclic Voltammetry (CV) scanning in the solution, and scanning for 20 sections in a voltage range of-0.4V-1.2V; after the scanning is completedWashing with water, and air drying to obtain activated SPCE.
(2) And (3) placing the activated SPCE electrode into 5mL of 0.01% chloroauric acid solution, depositing for 120s at a constant potential of-0.5V, washing for 3 times by using pure water after deposition is finished, and drying by blowing to obtain the Au NPs/SPCE electrode. And soaking the Au NPs/SPCE electrode in 2.5% glutaraldehyde for 15 min, washing with PBS (pH 7.0) for 3 times, drying, then dropwise adding 5 mu L H-rGO-Pt @ Pd NPs suspension, incubating for 30min, washing with PBS for 3 times, and drying to obtain H-rGO-Pt @ Pd NPs/Au NPs/SPCE. Dripping 2 mu L of carboxylated GPC3 aptamer onto a H-rGO-Pt @ Pd NPs/Au NPs/SPCE sensing interface, incubating for 1H, washing the aptamer which cannot be fixed on the interface, dripping 5 mu L of 0.5% BSA solution for sealing, and naturally drying to obtain GPC3Aptthe/H-rGO-Pt @ Pd NPs/Au NPs/SPCE sensing interface. Stored in a 4 ℃ refrigerator prior to use.
3. GPC3 Standard Curve
(1) Dropwise adding standard GPC3 solution to GPC3 obtained in step 2AptAnd putting the/H-rGO-Pt @ Pd NPs/Au NPs/SPCE sensing interface into an incubator at 25 ℃ for incubation for 1H, and washing the sensing interface with a PBS solution to obtain the GPC3 electrochemical biosensor.
(2) The working electrode obtained above was then placed in a PBS support solution (0.2 mol/L, pH 6.0) and the peak current was recorded using a DPV scan from the electrochemical workstation. When the concentration of GPC3 is in the range of 0.001-10 [ mu ] g/mL, the response current increases along with the increase of the concentration of GPC3, the DPV curve is shown in figure 3, the relation between the sensor current response value (Y) and the concentration (X) of GPC3 is linear, the standard curve is shown in figure 4, the linear regression equation is Y =1.7814+1.3576X, and the correlation coefficient is 0.9963. Three times the standard deviation of the blank was defined as the lower detection limit and the lowest detection limit of GPC3 was calculated to be 0.1845 ng/mL.
4. Detection of actual samples
mu.L of a known concentration of GPC3 solution (1. mu.g/mL, 5. mu.g/mL, 10. mu.g/mL) was added dropwise to GPC3Aptthe/H-rGO-Pt @ PdNPs/Au NPs/SPCE sensing interface, while 100. mu.L of human serum sample was added to 5mL of PBS support solution (0.2 mol/L, pH 6.0). And (4) placing the working electrode in the PBS supporting solution for DPV scanning according to the step 3, and recording the current value.According to the standard curve Y =1.7814+1.3576X in the step 3, the concentration of the GPC3 solution in the corresponding actual sample is calculated, and the detection result is shown in Table 1.
TABLE 1 results of GPC3 detection in actual serum samples (test (n = 3))
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not intended to limit the present invention in any way, and all simple modifications, equivalent variations and modifications made to the above embodiments according to the technical spirit of the present invention are within the scope of the present invention.
Claims (8)
1. A method for detecting GPC3 based on H-rGO-Pt @ Pd NPs nano composite material comprises the following steps:
step 1: preparation of H-rGO-Pt @ Pd NPs material
(1) Preparing reducing graphene oxide: pouring graphene oxide into distilled water, performing ultrasonic treatment by using an ultrasonic cell disruption instrument, and dissolving uniformly to prepare a GO aqueous solution; adding ascorbic acid into the GO aqueous solution to reduce GO to obtain RGO;
(2) preparing heme/reduced graphene oxide: adding heme into ammonia water to dissolve to obtain heme solution; mixing the heme solution with the RGO solution, mixing with a hydrazine hydrate solution, and stirring to obtain an H-rGO mixed solution;
(3) preparing a heme/reduced graphene oxide/platinum @ palladium composite material: adding PDDA and NaCl into the H-rGO solution, and stirring for reaction to obtain a PDDA modified H-rGO solution; mixing Na2PtCl6And Na2PdCl4Adding the solution into a PDDA modified H-rGO solution, stirring for reaction, then adding an ethylene glycol solution into the solution for mixing, adjusting the pH value of the mixed solution by using NaOH, and carrying out reflux reaction; centrifugally cleaning to obtain the H-rGO-Pt @ Pd NPs composite nano material;
step 2: electrode modification and biosensing interface construction
(1) Placing the screen-printed electrode in H2SO4In the solution, cyclic voltammetry scanning is carried out to obtain an activated screen printing electrode, and the screen printing electrode is washed clean by water;
(2) placing the activated screen printing electrode into a chloroauric acid solution, carrying out constant potential deposition, and washing the electrode clean with water after the deposition is finished to obtain an Au NPs/SPCE electrode;
(3) soaking the Au NPs/SPCE electrode with glutaraldehyde, washing with PBS, drying, then dropwise adding H-rGO-Pt @ Pd NPs suspension for incubation for a period of time, washing with PBS, and drying in the air to obtain the H-rGO-Pt @ Pd NPs/Au NPs/SPCE electrode;
(4) dropping the carboxylated GPC3 aptamer to the sensor interface, incubating for a period of time, washing the GPC3 aptamer which is not fixed to the interface with PBS solution, dropping bovine serum albumin solution for sealing to obtain GPC3Aptthe/H-rGO-Pt @ Pd NPs/AuNPs/SPCE sensing interface is dried for standby;
and step 3: GPC3 Standard Curve
(1) Dropwise adding standard GPC3 solution to GPC3 obtained in step 2AptIncubating a sensing interface of/H-rGO-Pt @ Pd NPs/Au NPs/SPCE for a period of time, washing the sensing interface with a PBS solution to obtain a working electrode, and airing the working electrode for later use;
(2) putting the working electrode into a PBS solution, adopting DPV scanning of an electrochemical workstation, and recording the peak current of the working electrode;
(3) detecting GPC3 with different concentrations, drawing a standard curve, and calculating the lowest detection limit of the method;
and 4, step 4: detection of actual samples
(1) GPC3 obtained in step 2Apta/H-rGO-Pt @ Pd NPs/Au NPs/SPCE sensing interface, dripping an actual sample to be detected, incubating for a period of time, cleaning with a PBS solution to obtain a working electrode, and airing for later use;
(2) putting the working electrode into a PBS solution, adopting DPV scanning of an electrochemical workstation, and recording the peak current of the working electrode;
(3) and (4) obtaining the concentration of GPC3 in the actual sample to be tested according to the standard curve in the step 3.
2. A method of detecting GPC3 according to claim 1, characterized in that: in step 1, the ammonia water solution is 10 mu L80%, the hydrazine hydrate solution is 8 mu L, and the PDDA is 2mL and 0.2%.
3. A method of detecting GPC3 according to claim 1, characterized in that: na described in step 12PtCl6And Na2PdCl4All are 2mL and 20 mmol/L.
4. A method of detecting GPC3 according to claim 1, characterized in that: in step 1, the NaCl is 5mL and 0.2mol/L, the NaOH is 1mol/L, and the EG is 10 mL.
5. A method of detecting GPC3 according to claim 1, characterized in that: placing the electrode in H as described in step 22SO4After cyclic voltammetry scanning, the electrode is washed clean by pure water, then is put into chloroauric acid solution to be respectively subjected to cyclic voltammetry scanning, and finally is washed by pure water and dried for standby.
6. A method of detecting GPC3 according to claim 1, characterized in that: the deposition solution for the nano-gold in the step 2 is chloroplatinic acid with the concentration of 0.01 percent, the deposition condition is-0.5V, and the deposition time is 120 s.
7. A method of detecting GPC3 according to claim 1, characterized in that: in step 2, the concentration of the BSA solution is 0.5%, the concentration of PBS is 0.2mol/L, the pH value is 7.0, and the concentration of GPC3 aptamer is 5 mu mol/L.
8. A method of detecting GPC3 according to claim 1, characterized in that: the incubation temperature of the electrode in the step 3 and the step 4 is 25 ℃, the incubation time is 1 hour, the linear scanning range is-0.4V-1.2V, and the scanning speed is 0.05V/s.
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| CN113203780A (en) * | 2021-05-13 | 2021-08-03 | 桂林电子科技大学 | Method for detecting GPC3 by using label-free aptamer sensor |
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| CN113203859A (en) * | 2021-05-13 | 2021-08-03 | 桂林电子科技大学 | Method for visually detecting GPC3 based on H-rGO-Pt @ Pd NPs nanoenzyme |
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| CN114965637A (en) * | 2022-04-24 | 2022-08-30 | 桂林电子科技大学 | Method for constructing sandwich type aptamer sensor based on nanocomposite to detect GPC3 |
| CN114965637B (en) * | 2022-04-24 | 2023-08-18 | 桂林电子科技大学 | Method for detecting GPC3 by constructing sandwich aptamer sensor based on nanocomposite |
| CN115032252A (en) * | 2022-04-28 | 2022-09-09 | 江南大学 | Electrochemical sensing analysis method for detecting ochratoxin A |
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