CN110146581A - A method of alpha-fetoprotein is detected based on RGO-CS-Fc/Au NPs nanocomposite combination aptamers - Google Patents
A method of alpha-fetoprotein is detected based on RGO-CS-Fc/Au NPs nanocomposite combination aptamers Download PDFInfo
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Abstract
A method of alpha-fetoprotein being detected based on RGO-CS-Fc/Au NPs nanocomposite combination aptamers, is modified RGO-CS-Fc/Au NPs on screen printing electrode surface using electro-deposition techniques and electrostatic adsorption.AFP aptamers are supported on RGO-CS-Fc/Au NPs material surface by nanotechnology and intermolecular force, there are composite material surfaces in the form of single-stranded structure due to its unstable space structure for aptamers.After AFP is added in electrode surface, AFP can be specifically bound with AFP aptamers, generate stable space structure, be arranged in electrode surface so as to orderly.Current value is detected by DPV method, and depicts the relation curve of the electric current Yu alpha-fetoprotein concentration, realizes the quantitative detection to alpha-fetoprotein.This method is easy to operate, time saving, expense is low and has lower detection limit.
Description
Technical field
The invention belongs to field of biological detection, and in particular to one kind detects first based on nanocomposite combination aptamers
The method of fetoprotein.
Background technique
Alpha-fetoprotein (AFP) is to belong to one of albumin family acidoglycoprotein, it is residual by more than 500 a amino acid
The single polypeptide chain that base is formed.For normal adult, AFP is extremely low in the intracorporal content of people.AFP detection method is mainly put
Radioimmunoassay method, fluoroimmunoassay, enzyme linked immunosorbent assay, chemiluminescence immunoassay, streaming immunoassay,
Electrochemical immunosensor, piezoelectric immunosensor and polypeptide compound protein microarray etc..104677889 B's of publication number CN
Patent of invention is related to a kind of method of magnetic immuno probe in detecting alpha-fetoprotein based on luminol functionalization.Utilize luminol
The magnetic microsphere labelled AFP secondary antibody of functionalization, so that the magnetic immuno probe of luminol functionalization be made.In gold electrode
Surface constructs " alpha-fetoprotein primary antibody/alpha-fetoprotein antigen/luminol functionalization magnetic immuno probe " formula composite construction, passes through
Electrochemiluminescence signal realization is detected to sensitive, the specific detection of alpha-fetoprotein.The hair of 105823886 B of publication number CN
Bright patent is related to a kind of method that ferrocene dicarboxylic acid/platinum nanoparticle/DNA enzymatic compound prepares and its detects alpha-fetoprotein.Shen
Please number be CN104569435B patent of invention, disclose a kind of preparation of unmarked optical electro-chemistry alpha-fetoprotein immunosensor
Method, in titania nanoparticles substrate, using optical electro-chemistry synthetic method, three metals for preparing dendrite nano bar-shape are closed
Gold nano-material, and then the unmarked optical electro-chemistry immunosensor of detection alpha-fetoprotein has been made.These method instruments
Expensive, complicated for operation, time-consuming and technical requirements are high, need to establish a kind of quick, sensitive, easy to operate AFP detection method.
Summary of the invention
Technical problem to be solved by the invention is to provide one kind based on reproducibility graphene oxide-chitosan-ferrocene/
The nanocomposite combination aptamers of nanogold (RGO-CS-Fc/Au NPs) improve sensitive come the method for detecting alpha-fetoprotein
Degree, enhancing specificity.
In order to solve the technical problem, made of electro-deposition techniques and electrostatic adsorption based on RGO-CS-Fc/
The AFP nanometer aptamers electrochemica biological sensor of Au NPs.Using graphene and nano metal to the high load of AFP aptamers
Ability and good electron transmission effect and AFP aptamers act on the specific recognition of AFP, using electrochemical workstation
Differential Pulse Voltammetry (DPV), record its peak current.It is suitable to the incubation temperature of AFP, incubation time, the pH value of PBS and AFP
Ligand concentration is optimized, and depicts standard curve, obtains accurate AFP concentration by comparing with standard working curve.With
Existing method is compared, and operation is relatively easy, and specificity is high, and the consumption of time and expense is less, can reach 1.013 ng/mL's
Detection limit.
Testing principle of the invention are as follows: repaired RGO-CS-Fc/Au NPs using electro-deposition techniques and electrostatic adsorption
Decorations are on screen printing electrode surface.AFP aptamers are supported on RGO-CS-Fc/ by nanotechnology and intermolecular force
Au NPs material surface, there are bio-sensing interfaces in the form of single-stranded structure due to its unstable space structure for aptamers.
After AFP is added on bio-sensing interface, AFP can be specifically bound with AFP aptamers, generate stable space structure, from
And orderly it can be arranged in electrode surface.By DPV method detect AFP before and after PBS solution (0.2 mol/L,
PH6.0 the electrochemical signals (sweep speed 0.01V/s, the voltage range of scanning are -0.4V-1.2V) in), and depict
The relation curve of the electric current and AFP concentration, to realize the detection to AFP.
The present invention follows the steps below:
The preparation of step 1:RGO-CS-Fc material
(1) preparation of reproducibility graphene oxide (RGO): graphene oxide (GO) pours into distilled water, broken using ultrasonic cell
Broken instrument ultrasound, is sufficiently uniformly dissolved it, GO aqueous solution is made.It takes above-mentioned GO aqueous solution to be placed in a beaker, ascorbic acid is added
(AA) so that it is restored GO, obtain RGO.
(2) preparation of chitosan-ferrocene (CS-Fc): chitosan (CS) is added in acetic acid solution, it is molten to obtain chitosan
Liquid.Ferrocenecarboxylic acid (Fc) is mixed with above-mentioned chitosan solution, with carbodiimide/n-hydroxysuccinimide (EDC/NHS)
Activation, stirs to obtain CS-Fc compound.
(3) reproducibility graphene oxide-chitosan-ferrocene (RGO-CS-Fc) composite material preparation: RGO suspension is taken
It is added in CS-Fc solution, EDC/NHS activation is centrifugated to obtain RGO-CS-Fc suspension.
Step 2: the modification of electrode and the building at bio-sensing interface
(1) screen printing electrode (SPE) is placed in H2SO4In solution, cyclic voltammetry scan, the screen printing after being activated are carried out
Brush electrode is rinsed with water clean.
(2) by the screen printing electrode merging chlorauric acid solution after activation, potentiostatic electrodeposition is carried out, is used after deposition
Water is clean by electrode washing, obtains Au NPs/SPE electrode.
(3) Au NPs/SPE electrode is impregnated with glutaraldehyde, is washed with PBS, dried up, it is suspended that RGO-CS-Fc is then added dropwise
Liquid is incubated for a period of time, and PBS washing dries, obtains RGO-CS-Fc/Au NPs/SPE electrode.
(4) it takes amidized AFP aptamers (AFP aptamer) to be added drop-wise to sensor interface, is incubated for a period of time, use
PBS solution washs the AFP aptamers for being not attached to interface, and bovine serum albumin (BSA) solution is added dropwise and is closed, AFP is obtained
Aptamer/RGO-CS-Fc/Au NPs/SPE sensing interface, dries spare.
Step 3: the Specification Curve of Increasing of alpha-fetoprotein
(1) standard AFP solution is added drop-wise to AFP aptamer/RGO-CS-Fc/Au NPs/SPE that step 2 obtains and senses boundary
Face is incubated for a period of time, is cleaned with PBS solution, obtain working electrode, dried spare.
(2) working electrode is put into PBS solution, is scanned using the DPV of electrochemical workstation, records its peak current.
(3) alpha-fetoprotein of various concentration is detected respectively, draws standard curve, calculates the minimum inspection of this method
Survey limit.
Step 4: the detection of AFP in actual sample
(1) practical sample to be measured is added dropwise in the AFP aptamer/RGO-CS-Fc/Au NPs/SPE sensing interface obtained in step 2
Product are incubated for a period of time, are cleaned with PBS solution, obtain working electrode, dried spare.
(2) working electrode is put into PBS solution, is scanned using the DPV of electrochemical workstation, records its peak current.
(3) standard curve according to step 3 obtains the concentration of alpha-fetoprotein in the actual sample to be measured.
Further, acetic acid solution is 100mL 1% in the step 1.
Further, EDC/NHS concentration is 10 mmol/L in the step 1.
Further, the material prepared is mixed in the step 1, is activated with EDC/NHS, be centrifuged, obtain RGO-CS-Fc
Suspension.
Further, H in the step 22SO4Solution concentration is 0.5 mol/L.
Further, scanning voltage is 1.2 V of -0.4 V- in the step 2, and scanning number of segment is 20.
Further, electrode is placed in H in the step 22SO4It is clean with pure water rinsing after middle carry out cyclic voltammetry scan
Afterwards, then electrode is placed in chlorauric acid solution and carries out cyclic voltammetry scan respectively, finally dried with pure water rinsing spare.
Further, in the step 2, the gold chloride concentration used is 0.01%, and sedimentary condition is -0.5 V, sedimentation time
120s。
Further, in the step 2, glutaraldehyde concentration 2.5%.
Further, the concentration that the BSA solution concentration is 0.5%, PBS is 0.2 mol/L, pH value 6.0.
Further, it is 0.1 μm of ol/L that AFP, which is adapted to bulk concentration, in step 2.
Further, AFP aptamers are 37 °C in the incubation temperature of electrode, and incubation time is 3 hours.
It is preferred that the best incubation temperature of alpha-fetoprotein is 25 DEG C in the step 3, best incubation time is 30min.
It is preferred that -0.4 V-1.2 V of linear scan range in the step 3 and 4, sweep speed is 0.01 V/s.
Wherein, step 1 is that step 2 provides a kind of nanocomposite of high conductivity.Step 2 constitutes specific recognition first
The bio-sensing interface of fetoprotein, and be conducive to the transmitting of electronics.The building at bio-sensing interface is step 3 and step in step 2
Essential committed step in the Electrochemical Detection of alpha-fetoprotein in rapid 4.The working curve of the alpha-fetoprotein of step 3 is step
The measurement of AFP concentration provides calculation basis in 4 actual sample.It can be seen that step 1-4 is mutually supported, collective effect could be utilized
It is to identify that probe realizes the detection of alpha-fetoprotein with RGO-CS-Fc/Au NPs composite material and alpha-fetoprotein aptamers.
The invention has the following advantages over the prior art:
1, RGO-CS-Fc/Au NPs composite nano materials have large specific surface area, and electric conductivity is strong, can effectively improve detection
Rate；Wherein, the large specific surface area of graphene and nanogold, adsorption capacity are strong, AFP aptamers effectively can be fixed to electricity
Detectability is improved to guarantee the stability of sensor in the surface of pole；It is anti-that with AFP aptamers specific binding can occur for AFP
It answers, generates stable space structure.Compared with traditional sensor, not only volume is smaller for novel nano-material sensor, speed
Faster, and precision is higher, and reliability is higher.
2, using with AFP aptamers is to identify that the background interference of probe in detecting alpha-fetoprotein is small, can reach 1.013ng/mL
Detection limit.Affinity between aptamers and object is usually stronger than the affinity between antigen and antibody.In addition, aptamers
It is easier to be marked and modified by chemical method than antibody, these processing facilitate the functionalization of nanoparticle He its surface.
Detailed description of the invention
The schematic diagram of nanometer aptamer sensor detection AFP of the Fig. 1 based on RGO-CS-Fc/Au NPs；
Fig. 2 RGO(A) and transmission electron microscope picture RGO-CS-Fc(B)；
The scanning electron microscope phenogram of Fig. 3 electrode surface different modifying process；
The working curve of AFP nanometer aptamer sensor of the Fig. 4 based on RGO-CS-Fc/Au NPs；Fig. 4 A is that different AFP are dense
The DPV curve of degree, Fig. 4 B are the working curve of AFP aptamer sensor.
Specific embodiment
The following describes the present invention in detail with reference to the accompanying drawings and specific embodiments.
A method of alpha-fetoprotein, detection are detected based on RGO-CS-Fc/Au NPs nanocomposite combination aptamers
Principle is shown in Fig. 1.RGO-CS-Fc/Au NPs is modified in screen printing electrode using electro-deposition techniques and electrostatic adsorption
Surface.AFP aptamers are supported on RGO-CS-Fc/Au NPs material surface by nanotechnology and intermolecular force, are fitted
There are on bio-sensing interface in the form of single-stranded structure due to its unstable space structure for ligand.In bio-sensing interface
After AFP is added, AFP can be specifically bound with AFP aptamers, stable space structure be generated, so as to orderly arrangement
In electrode surface.By DPV method detect AFP before and after PBS solution (0.2 mol/L, pH6.0) in electrochemical signals
(sweep speed 0.01V/s, the voltage range of scanning are -0.4V-1.2V), and depict the relationship of the electric current Yu AFP concentration
Curve, to realize the detection to AFP.
Implementation steps are as follows:
1. the preparation of RGO-CS-Fc composite nano materials: weighing 5mg graphene oxide (GO), and GO is poured into 50 mL's
In distilled water, it is sufficiently uniformly dissolved it using ultrasonic cell disintegration instrument ultrasound 2h, the GO aqueous solution of 0.1 mg/mL is made.It takes
The above-mentioned GO aqueous solution of 10 mL is placed in a beaker, and 10mg ascorbic acid (AA), which is added, makes it restore GO, is placed in constant temperature digital display magnetic force and is added
12h is stirred on thermal agitation device persistently to get RGO；2 mg chitosans (CS) are added in the acetic acid solution of 100 mL 1%, use glass
Glass stick is stirred continuously uniformly, until observing bubble-free in solution, obtains 2.0 mg/mL CS solution of stable homogeneous.It weighs again
2 mg ferrocenecarboxylic acids (Fc) are mixed with above-mentioned 10 mL chitosan, with 10 mmol/L EDC/NHS activation, are stirred 24 h, are obtained
Chitosan-ferrocene compound；10 mL RGO suspensions are taken to be added in 10 mL ferrocene-chitosan (CS-Fc) solution, with dense
The EDC/NHS that degree is 10 mmol/L activates 30 min, and 20000 r/min centrifuge separation obtains RGO-CS-Fc suspension.It adopts
Phenetic analysis is carried out to RGO-CS-Fc with transmission electron microscope (TEM), is seen as shown in Figure 2.The TEM that Fig. 2A is RGO schemes, and RGO is in the form of sheets
Structure and very flat and smooth, partially folded place's fold.The TEM that Fig. 2 B is RGO-CS-Fc schemes, and sheet is also presented in RGO-CS-Fc
Structure, but RGO is dispersed into clearer fritter sheet by chitosan, occurs there are many more combined thereon due to ferrocene
Darker small pieces.
2. the pretreatment of electrode: screen printing electrode (SPE) is first soaked in 0.5mol/L H before use2SO4Solution
Middle progress cyclic voltammetric (CV) scanning, scans 20 sections in the voltage range of -0.4V -1.2V；Scanning is washed with water after completing
Only, it dries, the SPE activated.
3. the building of the modification of electrode and bio-sensing interface: the SPE electrode after activation is put into 4mL 0.01% chlorine gold
Acid solution is used pure water 3 times after the completion of 120 s of -0.5 V potentiostatic electrodeposition, deposition, and drying obtains Au NPs/SPE electricity
Pole.2.5% glutaraldehyde of Au NPs/SPE electrode is impregnated into 15 min, is washed 3 times with pH7.0 PBS, is dried up, 5 μ L are then added dropwise
RGO-CS-Fc suspension be incubated for 30 min, PBS wash 3 times, dry, obtain RGO-CS-Fc/Au NPs/SPE.Take 2 μ L amino
AFP aptamers (AP273, the 5'-GTGACGCTCCTAACGCTGACTCAGGTGCAGTTCTCGACTCGGTCTTGATGTGG of change
GT CCTGTCCGTCCGAACCAATC-NH2- 3') it is added dropwise on RGO-CS-Fc/Au NPs/SPE sensing interface, 3 h are incubated for,
Washing fails the aptamers fixed to interface, and the BSA solution that 6 μ L 0.5% are added dropwise is closed, and naturally dry obtains AFP
Aptamer/RGO-CS-Fc/Au NPs/SPE sensing interface.Using scanning electron microscope (SEM) to electrode surface different modifying process
It is characterized, is seen as shown in Figure 3.Fig. 3 A is that the SEM of bare electrode (SPE) schemes, and naked screen printing electrode surface is because it is intrinsic
Carbon particle and show particle arranged in a uniform；Fig. 3 B is the SEM figure for depositing the electrode (Au NPs/SPE) after Jenner's grain of rice, by
In the presence of gold nano grain, it can be seen that be uniform-distribution with the spherical particle of many brilliant whites, explanation on the surface layer of carbon particle
Gold nanoparticle Au NPs successful deposition has arrived electrode surface；Fig. 3 C is the electrode modified after RGO-CS-Fc composite nano materials
SEM figure, RGO-CS-Fc is nanoscale nanocomposite, and becoming gold surface, gap is smaller, and electrode surface is obviously dimmed,
And more one layer of black cover；In Fig. 3 D it can be seen that composite material surface cover thin film, it is known that AFP aptamers at
Function is fixed on electrode surface.
4. the drafting of alpha-fetoprotein standard curve: in AFP aptamer/RGO-CS-Fc/Au NPs/SPE sensing interface
2 μ L alpha-fetoprotein solution are added dropwise, is incubated for 30min at a temperature of 25 DEG C, is cleaned with 7.0 PBS solution of pH and distilled water, dries up, obtains
To working electrode.Fig. 3 E is the SEM figure that AFP is adsorbed on bio-sensing interface, comparison diagram 3D, it is known that AFP and AFP aptamers are special
Property combine after with neat structural arrangement in electrode surface.Then above-mentioned resulting working electrode is put into PBS and supports liquid
In (0.2 mol/L, pH6.0), is scanned using the DPV of electrochemical workstation, record its peak current.The DPV of different AFP concentration
Curve graph is shown in Fig. 3 A.When AFP concentration is within the scope of 0.001 ~ 10 μ g/mL, sensor current response (Y) and AFP concentration (X)
Between relationship it is linear, standard curve is shown in Fig. 3 B, and equation of linear regression is Y=7.2106+2.8699X, and related coefficient is
0.9856.The three times standard deviation of blank control is defined as Monitoring lower-cut, the lowest detection for calculating alpha-fetoprotein is limited to
1.013ng/mL。
5. the detection of AFP in practical serum sample: by AFP solution (the 1 μ g/mL, 5 μ g/mL, 10 μ of 2 μ L known concentrations
G/mL it) is added dropwise and adds in AFP aptamer/ RGO-CS-Fc/Au NPs/SPE electrode surface, while by 100 μ L human serum samples
Enter to 5 mL PBS and supports in solution (0.2 mol/L, pH6.0).According to described in step 4, working electrode is placed in above-mentioned
PBS supports to carry out DPV scanning, record current value in solution.According to standard curve Y=7.2106+2.8699X of step 4, calculate
The concentration of AFP solution in corresponding actual sample can be obtained, testing result is shown in Table 1.
The testing result of AFP in the practical serum sample of table 1
(note: the concentration of AFP uses chemistry hair by the 9th 24 hospital, joint logistics system army, the Chinese People's Liberation Army in serum sample
Light immunization measurement).
The above is only not to make limit in any form to the present invention to better embodiment of the invention
System, any simple modification that embodiment of above is made according to the technical essence of the invention, equivalent variations and modification,
Belong in the range of technical solution of the present invention.
Claims (9)
1. one kind is based on RGO-CS-Fc/Au NPs nanocomposite combination aptamers as identification probe in detecting alpha-fetoprotein
Method, sequentially include the following steps:
The preparation of step 1:RGO-CS-Fc material
(1) preparation of RGO: GO is poured into distilled water, using ultrasonic cell disintegration instrument ultrasound, is sufficiently uniformly dissolved it, is made
At GO aqueous solution；Above-mentioned GO aqueous solution is taken, ascorbic acid is added, so that it is restored GO, obtains RGO；
(2) preparation of CS-Fc: CS is added in acetic acid solution, chitosan solution is obtained；Fc is mixed with above-mentioned chitosan solution,
It is activated with EDC/NHS, stirs to obtain CS-Fc compound；
(3) preparation of RGO-CS-Fc composite material: taking RGO suspension to be added in CS-Fc solution, EDC/NHS activation, centrifugation point
From RGO-CS-Fc suspension；
Step 2: the modification of electrode and the building at bio-sensing interface
(1) screen printing electrode is placed in H2SO4In solution, cyclic voltammetry scan is carried out, the silk-screen printing electricity after being activated
Pole；
(2) potentiostatic electrodeposition in the screen printing electrode merging chlorauric acid solution after activation, will be carried out, it will with water after deposition
Electrode washing is clean, obtains Au NPs/SPE electrode；
(3) Au NPs/SPE electrode is impregnated with glutaraldehyde, is washed with PBS, dried up；RGO-CS-Fc suspension is added dropwise to be incubated for,
PBS washing, dries, obtains RGO-CS-Fc/Au NPs/SPE electrode；
(4) it takes amidized AFP aptamer to be added drop-wise to sensor interface, is incubated for a period of time, is washed with PBS solution unlocked
To the AFP aptamers at interface, bovine serum albumen solution is added dropwise and is closed, AFP aptamer/RGO-CS-Fc/Au is obtained
NPs/SPE sensing interface dries spare；
Step 3: the working curve of alpha-fetoprotein is drawn
(1) standard AFP solution is added drop-wise to AFP aptamer/RGO-CS-Fc/Au NPs/SPE that step 2 obtains and senses boundary
Face is incubated for, and PBS solution cleaning obtains working electrode, dries spare；
(2) working electrode is put into PBS solution, is scanned using the DPV of electrochemical workstation, records its peak current；
(3) alpha-fetoprotein of various concentration is detected respectively, draws standard curve, calculates the lowest detection of this method
Limit；
Step 4: the detection of AFP in actual sample
(1) practical sample to be measured is added dropwise in the AFP aptamer/RGO-CS-Fc/Au NPs/SPE sensing interface obtained in step 2
Product are incubated for, are cleaned with PBS solution, obtain working electrode, dried spare；
(2) working electrode is put into PBS solution, is scanned using the DPV of electrochemical workstation, records its peak current；
(3) standard curve according to step 3 obtains the concentration of alpha-fetoprotein in the actual sample to be measured.
2. according to the method for detecting alpha-fetoprotein described in claim 1, it is characterised in that: acetic acid solution quality described in step 1
Concentration is 1%, volume 100mL.
3. according to the method for detecting alpha-fetoprotein described in claim 1, it is characterised in that: EDC/NHS concentration described in step 1 is
10 mmol/L。
4. according to the method for detecting alpha-fetoprotein described in claim 1, it is characterised in that: be placed in electrode described in step 2
H2SO4After middle carry out cyclic voltammetry scan, with pure water rinsing it is clean after, then by electrode be placed in 0.01% chlorauric acid solution of 4mL into
Row potentiostatic electrodeposition is finally dried with pure water rinsing spare.
5. according to the method for detecting alpha-fetoprotein described in claim 1, it is characterised in that: for nanogold described in step 2
Deposition solution is the chloroplatinic acid that concentration is 0.01%, and sedimentary condition is -0.5 V, 120 s of sedimentation time.
6. according to the method for detecting alpha-fetoprotein described in claim 1, it is characterised in that: BSA solution described in step 2 is 6 μ L
0.5% BSA solution.
7. according to the method for detecting alpha-fetoprotein described in claim 1, it is characterised in that: electrode described in step 3 and step 4
Incubation temperature is 25 °C, and incubation time is 30 minutes.
8. according to the method for detecting alpha-fetoprotein described in claim 1, it is characterised in that: DPV described in step 3 and step 4 is swept
Retouching solution used is the PBS solution that pH value is 6.0.
9. according to the method for detecting alpha-fetoprotein described in claim 1, it is characterised in that: scanning described in step 3 and step 4
Range is -0.4V ~ 1.2V, sweep speed 0.01V/s.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111413384A (en) * | 2020-04-26 | 2020-07-14 | 桂林电子科技大学 | Method for detecting GPC3 based on RGO-CS-Hemin/Au NPs nano composite material |
| CN111413385A (en) * | 2020-04-26 | 2020-07-14 | 桂林电子科技大学 | Method for detecting GPC3 based on RGO-CS-Fc/Pt-Pd NPs nano composite material |
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| CN111413384A (en) * | 2020-04-26 | 2020-07-14 | 桂林电子科技大学 | Method for detecting GPC3 based on RGO-CS-Hemin/Au NPs nano composite material |
| CN111413385A (en) * | 2020-04-26 | 2020-07-14 | 桂林电子科技大学 | Method for detecting GPC3 based on RGO-CS-Fc/Pt-Pd NPs nano composite material |
| CN111505077A (en) * | 2020-04-26 | 2020-08-07 | 桂林电子科技大学 | Method for detecting GPC3 based on RGO-Hemin/Au NPs nano composite material |
| CN111413385B (en) * | 2020-04-26 | 2023-09-19 | 桂林电子科技大学 | GPC3 detection method based on RGO-CS-Fc/Pt-Pd NPs nanocomposite |
| CN111413384B (en) * | 2020-04-26 | 2024-03-15 | 桂林电子科技大学 | GPC3 detection method based on RGO-CS-Hemin/Au NPs nanocomposite |
| CN111638326A (en) * | 2020-05-20 | 2020-09-08 | 江苏大学 | Enzyme-free polymer immunoprobe and preparation method and application thereof |
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| CN112763563A (en) * | 2021-02-03 | 2021-05-07 | 桂林电子科技大学 | Method for detecting 1, 5-anhydroglucitol based on composite material modified LAPS chip |
| CN112763563B (en) * | 2021-02-03 | 2022-11-29 | 桂林电子科技大学 | Method for detecting 1, 5-anhydroglucitol based on composite material modified LAPS chip |
| CN113009159A (en) * | 2021-02-26 | 2021-06-22 | 长沙市信励致和科技有限责任公司 | Human chorionic gonadotropin detection method based on human chorionic gonadotropin peptide aptamer |
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