CN115184431A - Preparation method and application of molecularly imprinted electrochemical sensor adopting double-signal probe strategy - Google Patents
Preparation method and application of molecularly imprinted electrochemical sensor adopting double-signal probe strategy Download PDFInfo
- Publication number
- CN115184431A CN115184431A CN202210903400.8A CN202210903400A CN115184431A CN 115184431 A CN115184431 A CN 115184431A CN 202210903400 A CN202210903400 A CN 202210903400A CN 115184431 A CN115184431 A CN 115184431A
- Authority
- CN
- China
- Prior art keywords
- aunps
- igg
- spce
- solution
- mwcnts
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000000523 sample Substances 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 229960003351 prussian blue Drugs 0.000 claims abstract description 90
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims abstract description 41
- 229940098773 bovine serum albumin Drugs 0.000 claims abstract description 41
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 30
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 claims abstract description 22
- 229910052737 gold Inorganic materials 0.000 claims abstract description 19
- 239000002105 nanoparticle Substances 0.000 claims abstract description 19
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 18
- 229910021389 graphene Inorganic materials 0.000 claims abstract description 12
- 239000010931 gold Substances 0.000 claims abstract description 9
- 238000007650 screen-printing Methods 0.000 claims abstract description 8
- 239000003480 eluent Substances 0.000 claims abstract description 6
- 230000000379 polymerizing effect Effects 0.000 claims abstract description 5
- 230000000295 complement effect Effects 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 88
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 21
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 17
- 238000002484 cyclic voltammetry Methods 0.000 claims description 15
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 14
- 238000001514 detection method Methods 0.000 claims description 13
- 239000002131 composite material Substances 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 11
- 229920002873 Polyethylenimine Polymers 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 239000002048 multi walled nanotube Substances 0.000 claims description 9
- 239000002253 acid Substances 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- -1 potassium ferricyanide Chemical compound 0.000 claims description 8
- 238000001318 differential pulse voltammogram Methods 0.000 claims description 7
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 7
- 239000001103 potassium chloride Substances 0.000 claims description 7
- 235000011164 potassium chloride Nutrition 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 230000000903 blocking effect Effects 0.000 claims description 6
- 239000008055 phosphate buffer solution Substances 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 5
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 5
- 239000012498 ultrapure water Substances 0.000 claims description 5
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims description 4
- 238000006116 polymerization reaction Methods 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 230000008021 deposition Effects 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 238000001132 ultrasonic dispersion Methods 0.000 claims description 3
- 238000007605 air drying Methods 0.000 claims description 2
- 239000011248 coating agent Substances 0.000 claims description 2
- 238000000576 coating method Methods 0.000 claims description 2
- 230000009977 dual effect Effects 0.000 claims 3
- 230000011664 signaling Effects 0.000 claims 3
- 230000005518 electrochemistry Effects 0.000 abstract description 2
- 229940027941 immunoglobulin g Drugs 0.000 description 87
- ANRHNWWPFJCPAZ-UHFFFAOYSA-M thionine Chemical compound [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 description 33
- DCYOBGZUOMKFPA-UHFFFAOYSA-N iron(2+);iron(3+);octadecacyanide Chemical compound [Fe+2].[Fe+2].[Fe+2].[Fe+3].[Fe+3].[Fe+3].[Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] DCYOBGZUOMKFPA-UHFFFAOYSA-N 0.000 description 26
- 239000013225 prussian blue Substances 0.000 description 26
- 229920000128 polypyrrole Polymers 0.000 description 13
- 239000010410 layer Substances 0.000 description 10
- 230000003647 oxidation Effects 0.000 description 9
- 238000007254 oxidation reaction Methods 0.000 description 9
- 230000008859 change Effects 0.000 description 8
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 230000008569 process Effects 0.000 description 5
- 108060003951 Immunoglobulin Proteins 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 102000018358 immunoglobulin Human genes 0.000 description 4
- 229920000642 polymer Polymers 0.000 description 4
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 4
- FGUUSXIOTUKUDN-IBGZPJMESA-N C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 Chemical compound C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 FGUUSXIOTUKUDN-IBGZPJMESA-N 0.000 description 3
- OUYCCCASQSFEME-MRVPVSSYSA-N D-tyrosine Chemical compound OC(=O)[C@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-MRVPVSSYSA-N 0.000 description 3
- 229930195709 D-tyrosine Natural products 0.000 description 3
- 102000001554 Hemoglobins Human genes 0.000 description 3
- 108010054147 Hemoglobins Proteins 0.000 description 3
- 229940072221 immunoglobulins Drugs 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000027756 respiratory electron transport chain Effects 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 229960004799 tryptophan Drugs 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- GNFTZDOKVXKIBK-UHFFFAOYSA-N 3-(2-methoxyethoxy)benzohydrazide Chemical compound COCCOC1=CC=CC(C(=O)NN)=C1 GNFTZDOKVXKIBK-UHFFFAOYSA-N 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000000835 electrochemical detection Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 239000002086 nanomaterial Substances 0.000 description 2
- 239000013047 polymeric layer Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000007704 transition Effects 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 208000029462 Immunodeficiency disease Diseases 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000008260 defense mechanism Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 230000007813 immunodeficiency Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 125000004433 nitrogen atom Chemical group N* 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229920000307 polymer substrate Polymers 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
- 238000011946 reduction process Methods 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Engineering & Computer Science (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Hematology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Nanotechnology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of analytical chemistry and electrochemistry, and discloses a preparation method and application of a molecular imprinting electrochemical sensor adopting a double-signal probe strategy. Fixing a first template molecule IgG to a Prussian blue @ gold nanoparticle/graphene oxide/silk-screen printing carbon electrode through a gold-sulfur bond to obtain a first template molecule IgG, fixing a second template molecule IgM to a multi-wall carbon nanotube-thionine-gold nanoparticle/bovine serum albumin/IgG/Prussian blue @ gold nanoparticle/graphene oxide/silk-screen printing carbon electrode through a gold-sulfur bond, electrochemically polymerizing pyrrole, eluting the IgG and the IgM by using an eluent to form an imprinting cavity complementary with the two template molecules in a three-dimensional space, and obtaining the molecularly imprinted electrochemical sensor. The molecularly imprinted electrochemical sensor can be used for simultaneously detecting IgG and IgM.
Description
Technical Field
The invention belongs to the technical field of analytical chemistry and electrochemistry, and particularly relates to a preparation method and application of a molecular imprinting electrochemical sensor adopting a double-signal probe strategy.
Background
Immunoglobulins (Ig) play an important role in many defense mechanisms of an organism against various potentially destructive objects, such as viruses or bacteria. Five common classes of immunoglobulins, i.e., immunoglobulins a, G, M, E, and D, have been reported to exist in the human circulatory system. Too high Ig levels can cause rheumatoid arthritis, liver disease and infectious diseases, but low Ig levels can also cause humoral immunodeficiency and metabolic diseases. Since 12 months 2019, igG and IgM have become important indicators for the assessment of early or infectious stages due to outbreaks of new corona epidemics. Enzyme-linked immunoassay has been commonly used for the detection of IgG and IgM, but has disadvantages of expensive instruments, complicated operation, and time-consuming assay. The electrochemical detection has the advantages of high sensitivity, low cost, simple operation and the like, thereby having great potential in the aspect of medical treatment.
The molecular imprinting electrochemical sensor combines a molecular imprinting technology with an electrochemical sensing technology, and adopts a Molecular Imprinting Polymer (MIP) as a specific molecular recognition element to enhance the selective recognition performance of the electrochemical sensor. The molecular imprinting electrochemical sensor has the advantages of high sensitivity, good selectivity, high analysis speed, easy miniaturization and the like, and is widely applied to the detection of multiple targets in serum. However, to date, studies of MIPs have been directed primarily to small molecules. Since proteins belong to biological macromolecules, which have relatively large molecular weights, complex surface morphologies, and variable molecular conformations, it is necessary to find a suitable method for molecular imprinting of proteins.
Disclosure of Invention
The invention provides a preparation method and application of a molecularly imprinted electrochemical sensor for simultaneously detecting IgG and IgM.
The technical scheme is as follows: in order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a molecular imprinting electrochemical sensor adopting a double-signal probe strategy can be used for simultaneously detecting IgG and IgM. A silk-Screen Printing Carbon Electrode (SPCE) is used as a substrate electrode, the SPCE is modified by Graphene Oxide (GO), prussian blue @ gold nanoparticles (PB @ AuNPs) are electrodeposited on the surface of the GO/SPCE in one step, immunoglobulin G (IgG) is used as a first template molecule to be modified on the surface of the PB @ AuNPs/GO/SPCE, and PB is a vector for indicating IgG signal change. The remaining binding sites were blocked with Bovine Serum Albumin (BSA). Preparing multi-walled carbon nano-tubes-thionine-gold nano-particles (MWCNTs-TH-AuNPs), modifying the multi-walled carbon nano-tubes-thionine-gold nano-particles on the surface of BSA/IgG/PB @ AuNPs/GO/SPCE, modifying immunoglobulin M (IgM) on the surface of the MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE as a second template molecule, and using TH as a vector for indicating IgM signal change. And then, electropolymerizing pyrrole (Py) to form a polymer layer to further embed IgG and IgM, and finally eluting IgG and IgM template molecules by using an eluent to form an imprinting cavity so as to successfully prepare the molecularly imprinted electrochemical sensor.
A molecular imprinting electrochemical sensor adopting a double-signal probe strategy is characterized in that a first template molecule IgG is fixed on Prussian blue @ gold nanoparticles/graphene oxide/silk-screen printing carbon electrode PB @ AuNPs/GO/SPCE through a gold-sulfur bond to obtain IgG/PB @ AuNPs/GO/SPCE, a second template molecule IgM is fixed on multi-walled carbon nano-tube-thionine-gold nanoparticles/bovine serum albumin/IgG/Prussian blue @ gold nanoparticles/graphene oxide/silk-screen printing carbon electrode MWs-TH-AuCNTs/BSA/IgG/PB @ AuNPs/GO/SPCE through a gold-sulfur bond to obtain IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE; and finally, electrochemically polymerizing pyrrole to obtain PPy/IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE, eluting IgG and IgM by using an eluent to form an imprinting cavity complementary with the two template molecules in a three-dimensional space, and thus obtaining the molecularly imprinted electrochemical sensor.
Further, the preparation method of the Prussian blue @ gold nanoparticles/graphene oxide/silk-screen printing carbon electrode PB @ AuNPs/GO/SPCE comprises the following steps: dissolving anhydrous ferric trichloride, potassium ferricyanide and potassium chloride in a hydrochloric acid solution, and then adding a chloroauric acid solution into the solution to obtain a PB @ AuNPs solution; transferring the GO solution to be dripped on the surface of the SPCE by using a liquid transfer gun, and drying under an infrared lamp to obtain GO/SPCE; scanning in the potential range of 0-1V by cyclic voltammetry, and electrodepositing a PB @ AuNPs layer on the surface of GO/SPCE to form the PB @ AuNPs/GO/SPCE.
Further, the preparation method of the multi-walled carbon nanotube-thionine-gold nanoparticle/bovine serum albumin/IgG/Prussian blue @ gold nanoparticle/graphene oxide/screen-printed carbon electrode MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE comprises the following steps:
dropping IgG solution on the surface of PB @ AuNPs/GO/SPCE, after air-drying, transferring BSA as a blocking liquid drop to be applied on the surface of PB @ AuNPs/GO/SPCE to obtain BSA/IgG/PB @ AuNPs/GO/SPCE; dissolving MWCNTs in a polyethyleneimine water solution, and performing ultrasonic dispersion; mixing a TH aqueous solution and the solution together, stirring vigorously, washing the solid matter obtained after centrifugal separation for a plurality of times by using ultrapure water after the stirring is finished, dispersing the solid matter in water to obtain a MWCNTs-TH solution, adding the MWCNTs-TH solution into an AuNPs solution, stirring uniformly, and centrifuging to obtain a solid, namely the MWCNTs-TH-AuNPs composite material; the MWCNTs-TH-AuNPs composite material is ultrasonically dispersed in water, and is coated on the surface of BSA/IgG/PB @ AuNPs/GO/SPCE by moving with a liquid-moving gun to obtain the MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE.
Further, the elution step is as follows: and transferring an acetic acid solution with the volume fraction of 10% and containing 10% of lauryl sodium sulfate by using a liquid transfer gun, dripping the acetic acid solution on the surface of PPy/IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE, eluting for 0.5-2.5 h, and removing the IgG and IgM as template molecules.
Further, when a PB @ AuNPs layer is electrodeposited on the GO/SPCE surface, the scanning rate of the cyclic voltammetry is 90-110 mV s –1 The number of scanning turns is 10-30 turns.
Further, a phosphate buffer solution of pyrrole is dripped on the surface of IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE, and the scanning rate of cyclic voltammetry is 25-125 mV s –1 Preferably 75mV s –1 And 3-15 circles of deposition are performed to form the PPy polymerization layer.
Further, the concentration of the IgG is 0.5 to 2.0 [ mu ] g mL –1 (ii) a The IgM concentration is 400-600 ng mL –1 。
Further, the concentration of the hydrochloric acid solution is 0.1mM; the concentration of the chloroauric acid solution was 0.2mM.
The invention also provides application of the molecular imprinting electrochemical sensor adopting the double-signal probe strategy in detection of IgG and IgM.
Further, the molecularly imprinted electrochemical sensor is incubated in a solution containing IgG and IgM for 1-6 h, and a differential pulse voltammogram is recorded in a potential range of-0.5V, so that IgG and IgM can be detected simultaneously. The invention also provides a preparation method of the molecular imprinting electrochemical sensor adopting the double-signal probe strategy, which comprises the following steps:
step a, preparing a PB @ AuNPs solution: weighing anhydrous ferric trichloride, potassium ferricyanide and potassium chloride, dissolving the anhydrous ferric trichloride, the potassium ferricyanide and the potassium chloride in 0.1M hydrochloric acid solution, and then adding 0.2mM chloroauric acid solution into the hydrochloric acid solution to obtain PB @ AuNPs solution;
step b, preparing PB @ AuNPs/GO/SPCE: transferring 10 mu L of GO solution by using a liquid transfer gun, dripping the GO solution on the surface of SPCE, and drying under an infrared lamp to obtain GO/SPCE; scanning for 15 circles within a potential range of 0-1V by using a cyclic voltammetry at a certain scanning rate, and electrodepositing a PB @ AuNPs layer on the surface of GO/SPCE to form PB @ AuNPs/GO/SPCE;
step c, preparing BSA/IgG/PB @ AuNPs/GO/SPCE: firstly transferring 10 mu L of IgG solution with certain concentration by using a liquid transfer gun, dripping the IgG solution on the surface of PB @ AuNPs/GO/SPCE, drying at 4 ℃, then transferring 10 mu L of BSA as a blocking liquid drop, and coating the blocking liquid drop on the surface of the electrode to obtain BSA/IgG/PB @ AuNPs/GO/SPCE;
step d, preparing the MWCNTs-TH-AuNPs composite material: weighing 10mg of MWCNTs, dissolving in a Polyethyleneimine (PEI) water solution, and performing ultrasonic dispersion; mixing a TH aqueous solution with a certain concentration with the solution, stirring vigorously, after stirring is finished, washing solid substances obtained after centrifugal separation for a plurality of times by using ultrapure water, dispersing the solid substances in water to obtain an MWCNTs-TH solution, transferring 0.5mL of the MWCNTs-TH solution into 2.5mL of AuNPs solution by using a liquid transfer gun, stirring for a certain time, and centrifuging to obtain solid substances, namely the MWCNTs-TH-AuNPs composite material.
Step e, preparing IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE: ultrasonically dispersing the MWCNTs-TH-AuNPs composite material prepared in the step d into 4mL of water, transferring 10 mu L of the IgM solution with a liquid transfer gun to be dripped on the surface of the electrode in the step c, naturally drying the IgM solution, transferring 10 mu L of the IgM solution with a certain concentration to be dripped on the surface of the electrode, and drying the IgM solution/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE at 4 ℃;
step f, preparing PPy/IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE: transferring 100 mu L of 0.01M phosphate buffer solution containing 0.1M pyrrole by using a liquid transfer gun, dropwise adding the solution on the surface of the electrode prepared in the step e, and polymerizing for certain circles at a certain scanning rate within the potential range of-0.3-0.8V by using a cyclic voltammetry method to obtain PPy/IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE;
step g, preparing the molecularly imprinted electrochemical sensor: and (f) transferring 100 mu L of 10% (v/v) acetic acid solution containing 10% (w%) sodium dodecyl sulfate by using a liquid transfer gun, dripping the acetic acid solution on the surface of the electrode prepared in the step f, and eluting for a certain time to remove template molecules IgG and IgM, thereby obtaining the molecularly imprinted electrochemical sensor.
Further, in the step a, the mass ratio of anhydrous ferric trichloride, potassium ferricyanide and potassium chloride is 0.04-0.05: 0.08 to 0.09: 0.7-0.8, the volume ratio of 0.1M hydrochloric acid to 0.2mM chloroauric acid is 10-50: 0.6 to 0.7.
Further, in the step b, the volume of GO solution transferred by a liquid transfer gun is 8-12 mu L; the sweep rate of the cyclic voltammetry is 90-110 mV s –1 The number of scanning turns is 10-30 turns to modify the electrode, and a PB @ AuNPs layer can be formed on the surface of the GO/SPCE.
Further, in step c, the volume of IgG transferred by the liquid transfer gun is 8-12 mu L, and the concentration is 0.5-2.0 mu g mL –1 The volume of BSA blocking solution transferred by a pipetting gun is 10-30 mu L, and the incubation time is 0.5-2 h.
Further, in the step d, the mass of PEI is 5-15mg, the mass of MWCNTs is 5-15 mg, the volume of PEI aqueous solution for dissolving MWCNTs is 5-15 mL, the volume of TH solution transferred by a pipette is 0.5-1.5 mL, and the concentration is 1-3 mg mL –1 The volume of MWCNTs-TH is 0.25-1.0 mL, the volume of AuNPs solution is 1.5-3.0 mL, and the stirring time is 10-14 h. Wherein, the mass ratio of PEI to MWCNTsIs 5 to 15: 5-15, wherein the volume ratio of the PEI aqueous solution, the TH solution, the MWCNTs-TH solution and the AuNPs solution is 5-15: 0.5-1.5: 0.25 to 1.0:1.5 to 3.0.
Further, in the step e, the volume of MWCNTs-TH-AuNPs solution transferred by a liquid transfer gun is 8-12 muL, and the concentration of IgM transferred by the liquid transfer gun is 400-600 ng mL –1 。
Further, in step f, the volume of 0.01M phosphate buffer solution with pH 7.4 containing 0.1M pyrrole is removed and is 50-150 μ L, and the sweep rate of cyclic voltammetry is 25-125 mV s –1 Preferably 75mV s –1 The number of deposition turns is 3 to 15 turns, preferably 7 turns, forming a PPy polymeric layer.
Furthermore, in the step g, the volume of 10% acetic acid solution containing 10% sodium dodecyl sulfate by mass fraction transferred by a liquid transfer gun is 100 to 200 μ L, and the elution time is 0.5 to 2.5h, preferably 2h.
Further, the method comprises the following steps: the molecular imprinting electrochemical sensor is applied to IgG and IgM for simultaneous detection, the re-incubation time is 1-6 h, preferably 4h, and a differential pulse voltammogram is recorded in a potential range of-0.5V to realize the simultaneous detection of IgG and IgM.
A molecular imprinting electrochemical sensor adopting a double-signal probe strategy is prepared by the preparation method.
In a third aspect, the molecularly imprinted electrochemical sensor adopting the double-signal probe strategy is applied to the simultaneous detection of IgG and IgM.
In some embodiments, the application comprises: placing the molecularly imprinted electrochemical sensor in a solution containing 1 mu g mL – 1 IgG and 500ng mL –1 IgM solutions were incubated.
Further, the application further includes: recording a cyclic voltammogram of the molecularly imprinted sensor recombined with IgG and IgM in a potential range of-0.6-0.4V, recording a differential pulse voltammogram of the molecularly imprinted sensor recombined with IgG and IgM in a potential range of-0.5V, and realizing the simultaneous detection of IgG and IgM through the change of peak currents before and after recombining IgG and IgM through PB and TH.
Advantageous effects
The surface molecular imprinting is an effective method for synthesizing the protein imprinted polymer, can improve the mass transfer process of protein molecules and avoid residues of the protein molecules, and can also obtain binding sites with higher affinity in a polymer matrix, thereby improving the recognition efficiency. Electropolymerization is a simple method that can be used to form a western blotting substrate, and polypyrrole (PPy) is often used as a polymer substrate material for western blotting due to its high stability, biocompatibility and conductivity, and controllable thickness.
The nano materials such as gold nanoparticles (AuNPs), multiwalled carbon nanotubes (MWCNTs), graphene Oxide (GO) and the like have excellent characteristics of large specific surface area, strong conductivity, high biocompatibility and the like, are often used for modifying electrodes, and can remarkably improve the analysis performance of an electrochemical sensor, such as high sensitivity and high electron transfer rate. When the nano materials are used for a molecular imprinting electrochemical sensor, abundant active sites can be provided for combining template molecules.
During electrochemical detection, when an electrochemical signal of an object to be detected is not obvious, the change of the concentration of the object to be detected is often indicated by an electrochemical vector, so that the detection of the concentration of the object to be detected is realized. Prussian Blue (PB) and Thionine (TH) are often selected as electrochemical mediators due to their good redox reversibility.
According to the preparation method and the application of the molecular imprinting electrochemical sensor adopting the double-signal probe strategy, igG and IgM template molecules in the polypyrrole layer are eluted, and then an imprinting cavity complementary to the IgG and IgM template molecules in a three-dimensional space is left, so that the imprinting cavity is favorable for electron transmission of PB and TH in a redox process. After the IgG and IgM molecules are recombined, the imprinting cavities are occupied again, so that the oxidation peak current of PB and TH is reduced, and the IgG and IgM can be detected simultaneously.
Drawings
FIG. 1 is a differential pulse voltammogram of SPCE, GO/SPCE, PB @ AuNPs/GO/SPCE, igG/PB @ AuNPs/GO/SPCE, BSA/IgG/PB @ AuNPs/GO/SPCE prepared in example I;
FIG. 2 is a differential pulse voltammogram of BSA/IgG/PB @ AuNPs/GO/SPCE, MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE, igM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE prepared in example one;
FIG. 3 is a cyclic voltammogram of a molecularly imprinted electrochemical sensor prepared in the first example after polymerizing pyrrole, eluting, and incubating in IgG and IgM solutions;
FIG. 4 is a differential pulse voltammogram of a molecularly imprinted electrochemical sensor prepared in the first example for detecting solutions containing different concentrations of IgG and IgM;
FIG. 5 is a graph showing the decrease of TH peak current (. DELTA.I) when the molecularly imprinted electrochemical sensor prepared in example one was used for simultaneous detection of IgG and IgM TH ) Linear dependence of IgG concentration logarithm (lgC) on (A) and reduction of PB peak current (. DELTA.I) PB ) A linear plot (B) against the log IgM concentration (lgC);
FIG. 6 is a diagram of the UV-Vis absorption spectrum of MWCNTs-TH-AuNPs prepared in the first example.
Fig. 7 shows the specificity of the molecularly imprinted electrochemical sensor prepared in example one.
Fig. 8 is a cyclic voltammogram of a non-imprinted electrochemical sensor prepared in comparative example one after polymerization of pyrrole, elution, and incubation in IgG and IgM solutions.
Detailed Description
The present invention will be further described with reference to the following drawings and specific examples, but the present invention is not limited to the following examples.
The first embodiment is as follows:
a preparation method of a molecular imprinting electrochemical sensor adopting a double-signal probe strategy comprises the following steps:
(1) 0.0405g anhydrous ferric chloride, 0.0823g potassium ferricyanide and 0.7455g potassium chloride are weighed and dissolved in 100mL of 0.1M hydrochloric acid solution, and then 0.689mL of 29mM chloroauric acid solution is added into the solution, thus obtaining PB @ AuNPs solution.
(2) Weighing 2mg of GO powder, dissolving the GO powder in 1mL of ultrapure water, uniformly dispersing by ultrasonic, dripping the GO powder on the surface of SPCE, and drying under an infrared lamp to obtain GO/SPCE. As in FIG. 1, GO/SPCE has no oxidation peak; pipetting with pipette 10 muL PB @ AuNPs solution is put on GO/SPCE, and 100mV s is used in the potential range of 0-1V by cyclic voltammetry –1 The scanning speed of (1) is that a PB @ AuNPs layer is electrodeposited on the surface of GO/SPCE to form the PB @ AuNPs/GO/SPCE. As shown in FIG. 1, PB @ AuNPs/SPCE showed oxidation peak of PB at 0.1V, which indicates that PB is an excellent electrochemical vector. The invention uses PB as a first electrochemical probe to indicate the change of signal of IgG.
(3) First, 10. Mu.L of 1.0. Mu.g mL was pipetted using a pipette gun –1 And (3) modifying the surface of the electrode obtained in the step (2) by using an IgG solution, and incubating at 4 ℃. As shown in FIG. 1, compared with PB @ AuNPs/SPCE, the oxidation peak current of IgG/PB @ AuNPs/GO/SPCE is significantly reduced, because IgG is a biological macromolecule with poor conductivity, and the electron transfer on the surface of the electrode is inhibited after the IgG is combined with AuNPs through sulfydryl; then 10. Mu.L BSA was taken as blocking solution to incubate the electrode at 4 ℃ for 1h, thus obtaining BSA/IgG/PB @ AuNPs/GO/SPCE, as shown in FIG. 1, the current continuously decreased due to the poor conductivity of BSA itself.
(4) And (4) transferring 10 mu L of MWCNTs-TH-AuNPs solution, dripping the solution on the surface of the electrode in the step (3), and drying at room temperature to obtain MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE. As shown in FIG. 2, the oxidation peak of TH appears at-0.2V in MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE, which shows that TH is also an excellent electrochemical vector, and the oxidation peak position of PB is almost unchanged. TH is used as a second electrochemical probe and indicates the signal change of IgM; pipette 10. Mu.L of 500ng mL with pipette –1 And (3) incubating the electrode with the IgM solution at 4 ℃ to obtain IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE. As shown in FIG. 2, the oxidation peak of TH is reduced, and the electron transport is hindered due to the combination of the IgM, a biomacromolecule having poor conductivity, with AuNPs among MWCNTs-TH-AuNPs through a thiol group.
(5) Transferring 100 μ L of 0.01M phosphate buffer solution containing 0.1M pyrrole, dripping on the surface of the electrode prepared in step (4), and performing cyclic voltammetry at a potential range of-0.3-0.8V in 75mV s –1 The scanning speed is 7 times of polymerization, and PPy/IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE can be obtained. As shown in FIG. 3, the peaks of PB and TH disappear after electropolymerization of polypyrrole, since the oxygen of PB and TH is blocked after the polypyrrole layer is formed on the electrode surfaceAnd (4) carrying out a reduction process.
(6) And (4) transferring 100 mu L of 10% acetic acid elution solution containing 10% sodium dodecyl sulfate to the surface of the electrode prepared in the step (5) for eluting the template molecules, and after eluting for 2 hours, washing the electrode by using ultrapure water and phosphate buffer solution for multiple times alternately to remove IgG and IgM remained on the surface, thereby obtaining the molecular imprinting electrochemical sensor. As shown in fig. 3, PB and TH redox peaks appear, and due to the fact that acidic eluent can break down protein structures, igG and IgM can be eluted from a polymeric layer, so that a blotting cavity is formed, and the redox reaction of PB and TH is facilitated. 10. Mu.L of the suspension containing 1.0. Mu.g mL of the reagent –1 IgG and 500ng mL – 1 Solutions of IgM were incubated with the molecularly imprinted electrochemical sensor for 4h at 4 ℃. As shown in fig. 3, the peak currents of PB and TH are significantly reduced because IgG and IgM re-occupy the blot cavity, thereby inhibiting the redox process of PB and TH.
(7) The prepared molecularly imprinted electrochemical sensor contains IgG (0.05 ng mL) –1 ～100ng mL –1 ) And IgM (0.05 ng mL) –1 ～100ng mL –1 ) Incubated for 4h. As shown in FIG. 4, the oxidation peak at TH, occurring at about-0.23V, is indicative of a change in IgM concentration, and the oxidation peak at PB, occurring at about 0.11V, is indicative of a change in IgG concentration. As the IgG and IgM concentrations gradually increase, the peak currents of PB and TH gradually decrease due to the occupation of the corresponding imprinted cavities by IgG and IgM, thereby inhibiting the transport of electrons generated during the redox process of PB and TH.
(8) When the prepared molecularly imprinted electrochemical sensor detects IgG and IgM simultaneously, the reduction value of peak current and the concentration logarithm value are in a linear relation. As shown in FIG. 5A, the linear equation of IgM is Δ I TH (μA)＝1.662lgC+4.813(R 2 = 0.9962) linear range of 0.05ng mL –1 ～100ng mL –1 In FIG. 5B, the linear equation for IgG is Δ I PB (μA)＝2.130lgC+9.802(R 2 = 0.9945), linear range 0.05ng mL –1 ～100ng mL –1 . According to the triple signal-to-noise ratio, the detection limit of IgM is calculated to be 28.61pg mL –1 The limit of IgG detection was 23.93pg mL –1 . The results prove that the molecularly imprinted electrochemical sensor can detect IgG and IgM simultaneously.
(9) And (3) transferring 0.5mL of MWCNTs-TH solution into 2.5mL of AuNPs solution, stirring for 12h, and dispersing the solid obtained by centrifugation into water to obtain the solution of the MWCNTs-TH-AuNPs composite material. As shown in fig. 6, curve a is the uv-vis spectrum of AuNPs, the characteristic absorption band of AuNPs appears at 519nm, indicating that AuNPs were successfully prepared, curve b is the uv-vis spectrum of MWCNT-TH, two characteristic absorption bands appear at 285 and 601nm, belonging to the pi-pi transition of the aromatic ring of TH and the N-pi transition of the C = N bond, respectively, indicating that TH molecules are non-covalently attached to the surface of MWCNTs by pi-pi stacking, curve C is the uv-vis spectrum of MWCNT-TH-AuNPs, similar to the spectrum of MWCNT-TH (curve b), but the absorption band at 601nm is blue-shifted to 597nm, probably due to the coordination of the N atom of TH with Au, and the disappearance of the absorption band at 519nm is attributed to the aggregation of AuNPs on MWCNT-TH. The above results indicate that MWCNT-TH-AuNPs composite materials have been successfully synthesized.
(10) By detecting 100ng mL –1 IgG and IgM, and 10-fold concentration (1. Mu.g mL) –1 ) Bovine Serum Albumin (BSA), hemoglobin (HGB), L-tryptophan (L-Trp) and D-tyrosine (D-Tyr) to investigate the specificity of the molecularly imprinted electrochemical sensor (FIG. 7). The test result shows that after the sensor is incubated in 10 times concentration BSA, HGB, L-Trp and D-Tyr solution for 4 hours, the Delta I of the sensor PB And Δ I TH The molecular imprinting electrochemical sensor is obviously lower than the value of detecting IgG and IgM, and shows that the molecular imprinting electrochemical sensor has good specificity.
Comparative example one:
to further demonstrate the successful fabrication of the molecularly imprinted electrochemical sensor, a non-imprinted electrochemical sensor was additionally fabricated, which was operated in the same manner as in example steps (1) to (6) except that no IgG or IgM template molecules were added. As in fig. 8, no redox peaks of PB and TH occurred due to the polypyrrole layer hindering the redox process of PB and TH. Due to the absence of IgG and IgM, when the electrode is eluted by acidic eluent, a blotting cavity favorable for electron transfer cannot be formed, so that oxidation reduction of PB and TH still does not occur on the eluted electrodeAnd (4) peak. 10. Mu.L of the suspension containing 1.0. Mu.g mL of the reagent –1 IgG and 500ng mL –1 IgM solution was incubated at 4 ℃ for 4h with the molecularly imprinted electrochemical sensor, as shown in FIG. 8, and the current signal was almost unchanged, indicating that no imprinted cavity was present. By comparing the molecularly imprinted electrochemical sensor with a non-imprinted electrochemical sensor, the molecularly imprinted electrochemical sensor is further proved to be capable of detecting IgG and IgM simultaneously.
Claims (10)
1. A molecular imprinting electrochemical sensor adopting a double-signal probe strategy is characterized in that a first template molecule IgG is fixed to Prussian blue @ gold nano-particle/graphene oxide/silk-screen printing carbon electrode PB @ AuNPs/GO/SPCE through a gold-sulfur bond to obtain IgG/PB @ AuNPs/GO/SPCE, a second template molecule IgM is fixed to a multi-wall carbon nano-tube-thionine-gold nano-particle/bovine serum albumin/IgG/Prussian blue @ gold nano-particle/graphene oxide/silk-screen printing carbon electrode MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE through a gold-sulfur bond to obtain IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/SPCE; and finally, electrochemically polymerizing pyrrole to obtain PPy/IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE, eluting IgG and IgM by using eluent to form an imprinting cavity complementary with the two template molecules in three-dimensional space, and thus obtaining the molecularly imprinted electrochemical sensor.
2. The preparation method of the molecularly imprinted electrochemical sensor adopting the dual-signal probe strategy as claimed in claim 1, wherein the preparation method of the Prussian blue @ gold nanoparticles/graphene oxide/screen-printed carbon electrode PB @ AuNPs/GO/SPCE comprises the following steps: dissolving anhydrous ferric trichloride, potassium ferricyanide and potassium chloride in a hydrochloric acid solution, and then adding a chloroauric acid solution into the solution to obtain a PB @ AuNPs solution; transferring the GO solution by using a liquid transfer gun, dripping the GO solution on the surface of the SPCE, and drying under an infrared lamp to obtain GO/SPCE; scanning in a potential range of 0-1V by using a cyclic voltammetry method, and electrodepositing a PB @ AuNPs layer on the surface of the GO/SPCE to form the PB @ AuNPs/GO/SPCE.
3. The method for preparing a molecularly imprinted electrochemical sensor with a dual-signal probe strategy according to claim 1, wherein the method for preparing the multi-walled carbon nanotube-thionine-gold nanoparticle/bovine serum albumin/IgG/Prussian blue @ gold nanoparticle/graphene oxide/screen-printed carbon electrode MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE comprises:
dropping IgG solution on the surface of PB @ AuNPs/GO/SPCE, air-drying, then transferring BSA as a blocking liquid to drop coat the BSA on the surface to obtain BSA/IgG/PB @ AuNPs/GO/SPCE; dissolving MWCNTs in a polyethyleneimine water solution, and performing ultrasonic dispersion; mixing a TH aqueous solution and the solution together, stirring vigorously, washing the solid matter obtained after centrifugal separation for a plurality of times by using ultrapure water after the stirring is finished, dispersing the solid matter in water to obtain a MWCNTs-TH solution, adding the MWCNTs-TH solution into an AuNPs solution, stirring uniformly, and centrifuging to obtain a solid, namely the MWCNTs-TH-AuNPs composite material; ultrasonically dispersing the MWCNTs-TH-AuNPs composite material in water, transferring the MWCNTs-TH-AuNPs composite material by using a transfer gun and coating the MWCNTs-TH-AuNPs/GO/SPCE surface with the MWCNTs-TH-AuNPs composite material to obtain the MWCNTs-TH-AuNPs composite material
MWCNTs-TH-AuNPs/BSA/IgG/PB@AuNPs/GO/SPCE。
4. The method for preparing a molecularly imprinted electrochemical sensor adopting the dual-signaling probe strategy as claimed in claim 1, wherein the elution step comprises: using a liquid-transferring gun to transfer 10 percent by volume acetic acid solution containing 10 percent by mass of sodium dodecyl sulfate to be coated on the solution drop by drop
PPy/IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE surface, eluting for 0.5-2.5 h, and removing template molecules IgG and IgM.
5. The preparation method of the molecularly imprinted electrochemical sensor adopting the dual-signal probe strategy as claimed in claim 2, wherein the scanning rate of cyclic voltammetry is 90-110 mV s when a PB @ AuNPs layer is electrodeposited on the GO/SPCE surface –1 The number of scanning circles is 10-30 circles; the phosphate buffer solution of pyrrole is dripped on the surface of IgM/MWCNTs-TH-AuNPs/BSA/IgG/PB @ AuNPs/GO/SPCE, and the scanning rate of the cyclic voltammetry is 25-125 mV s –1 And 3-15 circles of deposition are performed to form the PPy polymerization layer.
6. The method for preparing the molecularly imprinted electrochemical sensor adopting the dual-signal probe strategy as claimed in claim 1, wherein the IgG concentration is 0.5-2.0 μ g mL –1 (ii) a IgM concentration of 400-600 ng mL –1 。
7. The method for preparing a molecularly imprinted electrochemical sensor adopting a dual-signal probe strategy according to claim 1, wherein the mass ratio of anhydrous ferric trichloride to potassium ferricyanide to potassium chloride is 0.04-0.05: 0.08 to 0.09:0.7 to 0.8; the mass ratio of PEI to MWCNTs is 5-15: 5 to 15 percent; the volume ratio of the PEI aqueous solution to the TH solution to the MWCNTs-TH solution to the AuNPs solution is 5-15: 0.5 to 1.5:0.25 to 1.0:1.5 to 3.0.
8. The method for preparing a molecularly imprinted electrochemical sensor adopting the dual signaling probe strategy of claim 2, wherein the concentration of the hydrochloric acid solution is 0.1mM; the concentration of the chloroauric acid solution is 0.2mM; the volume ratio of the hydrochloric acid to the chloroauric acid is 10-50: 0.6 to 0.7.
9. The use of the molecularly imprinted electrochemical sensor of the dual signaling probe strategy of claim 1 for the detection of IgG and IgM.
10. The use of claim 9, wherein the molecularly imprinted electrochemical sensor is incubated in a solution containing IgG and IgM for 1-6 h, and differential pulse voltammogram is recorded in a potential range of-0.5V to achieve simultaneous detection of IgG and IgM.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210903400.8A CN115184431B (en) | 2022-07-26 | 2022-07-26 | Preparation method and application of molecular imprinting electrochemical sensor with double-signal probe strategy |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210903400.8A CN115184431B (en) | 2022-07-26 | 2022-07-26 | Preparation method and application of molecular imprinting electrochemical sensor with double-signal probe strategy |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115184431A true CN115184431A (en) | 2022-10-14 |
| CN115184431B CN115184431B (en) | 2024-03-12 |
Family
ID=83521346
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210903400.8A Active CN115184431B (en) | 2022-07-26 | 2022-07-26 | Preparation method and application of molecular imprinting electrochemical sensor with double-signal probe strategy |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115184431B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117288820A (en) * | 2023-09-26 | 2023-12-26 | 天津大学 | Three-way electrochemical sensing system and method based on Internet of things communication |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102718984A (en) * | 2012-06-28 | 2012-10-10 | 陕西师范大学 | Preparation method of ofloxacin and 17beta-estradiol double-template molecularly-imprinted composite microsphere |
| US20130306485A1 (en) * | 2011-02-04 | 2013-11-21 | Council Of Scientific & Industrial Research | Molecularly Imprinted Conducting Polymer Film Based Aqueous Amino Acid Sensors |
| CN106198667A (en) * | 2016-07-01 | 2016-12-07 | 肇庆学院 | A kind of molecular imprinting electrochemical sensor for detecting trace bisphenol-A |
| WO2017039345A1 (en) * | 2015-09-01 | 2017-03-09 | 부산대학교 산학협력단 | One-time-use amperometric detection method for simultaneously measuring hemoglobin and glycated hemoglobin |
| CN107179348A (en) * | 2017-06-21 | 2017-09-19 | 安徽师范大学 | A kind of double-template trace electrochemical sensor and its preparation method and application |
| CN110687183A (en) * | 2019-10-30 | 2020-01-14 | 安徽师范大学 | Double-signal-double-recognition imprinting electrochemical sensor, preparation method thereof and ovalbumin detection method |
| CN111944104A (en) * | 2020-08-11 | 2020-11-17 | 华南师范大学 | Porous double-template molecularly imprinted polymer microsphere for detecting dengue NS1 protein and application thereof |
-
2022
- 2022-07-26 CN CN202210903400.8A patent/CN115184431B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130306485A1 (en) * | 2011-02-04 | 2013-11-21 | Council Of Scientific & Industrial Research | Molecularly Imprinted Conducting Polymer Film Based Aqueous Amino Acid Sensors |
| CN102718984A (en) * | 2012-06-28 | 2012-10-10 | 陕西师范大学 | Preparation method of ofloxacin and 17beta-estradiol double-template molecularly-imprinted composite microsphere |
| WO2017039345A1 (en) * | 2015-09-01 | 2017-03-09 | 부산대학교 산학협력단 | One-time-use amperometric detection method for simultaneously measuring hemoglobin and glycated hemoglobin |
| CN106198667A (en) * | 2016-07-01 | 2016-12-07 | 肇庆学院 | A kind of molecular imprinting electrochemical sensor for detecting trace bisphenol-A |
| CN107179348A (en) * | 2017-06-21 | 2017-09-19 | 安徽师范大学 | A kind of double-template trace electrochemical sensor and its preparation method and application |
| CN110687183A (en) * | 2019-10-30 | 2020-01-14 | 安徽师范大学 | Double-signal-double-recognition imprinting electrochemical sensor, preparation method thereof and ovalbumin detection method |
| CN111944104A (en) * | 2020-08-11 | 2020-11-17 | 华南师范大学 | Porous double-template molecularly imprinted polymer microsphere for detecting dengue NS1 protein and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 盖青青;屈锋;: "牛血清白蛋白和溶菌酶为双模板蛋白质的表面印迹聚合物的制备及吸附性能", 色谱, no. 05 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117288820A (en) * | 2023-09-26 | 2023-12-26 | 天津大学 | Three-way electrochemical sensing system and method based on Internet of things communication |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115184431B (en) | 2024-03-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Nemiwal et al. | Enzyme immobilized nanomaterials as electrochemical biosensors for detection of biomolecules | |
| Yang et al. | Frontiers in highly sensitive molecularly imprinted electrochemical sensors: Challenges and strategies | |
| Chen et al. | A novel bienzyme glucose biosensor based on three-layer Au–Fe3O4@ SiO2 magnetic nanocomposite | |
| Shiddiky et al. | Application of ionic liquids in electrochemical sensing systems | |
| Chen et al. | Electrochemical immunosensor for carcinoembryonic antigen based on nanosilver-coated magnetic beads and gold-graphene nanolabels | |
| Shahrokhian et al. | Application of thionine-nafion supported on multi-walled carbon nanotube for preparation of a modified electrode in simultaneous voltammetric detection of dopamine and ascorbic acid | |
| Morrin et al. | An amperometric enzyme biosensor fabricated from polyaniline nanoparticles | |
| Roy et al. | Introduction of selectivity and specificity to graphene using an inimitable combination of molecular imprinting and nanotechnology | |
| Janczuk-Richter et al. | Recent applications of bacteriophage-based electrodes: A mini-review | |
| Han et al. | Electrochemical immunoassay for thyroxine detection using cascade catalysis as signal amplified enhancer and multi-functionalized magnetic graphene sphere as signal tag | |
| Prasad et al. | Quantum dots-multiwalled carbon nanotubes nanoconjugate-modified pencil graphite electrode for ultratrace analysis of hemoglobin in dilute human blood samples | |
| CN115184431B (en) | Preparation method and application of molecular imprinting electrochemical sensor with double-signal probe strategy | |
| CN109085225B (en) | Preparation method of western blot sensor with magnetic electrode modified by one-step deposition method | |
| Negahdary et al. | Electrochemical nanobiosensors equipped with peptides: a review | |
| Filip et al. | Immobilization of concanavalin A lectin on a reduced graphene oxide-thionine surface by glutaraldehyde crosslinking for the construction of an impedimetric biosensor | |
| CN115586229A (en) | MXene nano material-based ethyl glucuronide electrochemical biosensor and preparation method and detection method thereof | |
| Lu et al. | Determination of Uric Acid and Norepinephrine by Chitosan‐Multiwall Carbon Nanotube Modified Electrode | |
| Zakiyyah et al. | Screen-printed carbon electrode/natural silica-ceria nanocomposite for electrochemical aptasensor application | |
| Negahdary et al. | Recent advances in electrochemical nanobiosensors for cardiac biomarkers | |
| Behyar et al. | Sensing of amino acids: Critical role of nanomaterials for the efficient biomedical analysis | |
| Mousazadeh et al. | Recent advantages of mediator based chemically modified electrodes; powerful approach in electroanalytical chemistry | |
| Upadhyay et al. | A biosensor for hydrogen peroxide based on single walled carbon nanotube and metal oxide modified indium tin oxide electrode | |
| Low et al. | An electrochemical genosensing assay based on magnetic beads and gold nanoparticle-loaded latex microspheres for vibrio cholerae detection | |
| Li et al. | A non-enzymatic electrochemical biosensor based on SiO 2–Au nanoparticles for hemoglobin detection | |
| CN105866221A (en) | Catalytic reduction hemoglobin electrochemical sensor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |