WO2012081940A2 - Impression au moyen d'une électrode à phase membranaire et détection de biomolécule utilisant ladite impression - Google Patents

Impression au moyen d'une électrode à phase membranaire et détection de biomolécule utilisant ladite impression Download PDF

Info

Publication number
WO2012081940A2
WO2012081940A2 PCT/KR2011/009746 KR2011009746W WO2012081940A2 WO 2012081940 A2 WO2012081940 A2 WO 2012081940A2 KR 2011009746 W KR2011009746 W KR 2011009746W WO 2012081940 A2 WO2012081940 A2 WO 2012081940A2
Authority
WO
WIPO (PCT)
Prior art keywords
electrode
membrane
ligand
target
enzyme
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.)
Ceased
Application number
PCT/KR2011/009746
Other languages
English (en)
Korean (ko)
Other versions
WO2012081940A3 (fr
Inventor
김민곤
안준형
성윤주
신용범
정효암
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Korea Research Institute of Bioscience and Biotechnology KRIBB
Original Assignee
Korea Research Institute of Bioscience and Biotechnology KRIBB
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Korea Research Institute of Bioscience and Biotechnology KRIBB filed Critical Korea Research Institute of Bioscience and Biotechnology KRIBB
Priority to US13/994,348 priority Critical patent/US9465003B2/en
Priority claimed from KR1020110136334A external-priority patent/KR101335246B1/ko
Publication of WO2012081940A2 publication Critical patent/WO2012081940A2/fr
Publication of WO2012081940A3 publication Critical patent/WO2012081940A3/fr
Anticipated expiration legal-status Critical
Priority to US15/269,779 priority patent/US20170010234A1/en
Ceased legal-status Critical Current

Links

Images

Classifications

    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • G01N33/5438—Electrodes
    • 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

Definitions

  • the present invention relates to a method for measuring a biomaterial combining the filtration characteristics of the membrane and the signal measurement of the electrode by preparing an electrode on the filtration membrane by a screen printing method.
  • Technology for quantitative measurement of biological substances is an important technique for food, medicine, and diagnosis.
  • the rapid measurement of microorganisms is a very important technique for diagnosis of food poisoning bacteria, measurement of environmental harmful bacteria, measurement of infectious bacteria, diagnosis of pathogenic viruses, and the like.
  • colony, DNA probe, and immunoassay methods are widely used as a method for examining the presence and concentration of commonly used pathogenic substances (microorganisms, proteins, etc.) (Jay JM. Modern Food Microbiology, 1986, 3rd, ed., P 95, Van Nostrand Reinhold Co., New York; Tenover FC., DNA Probes for Infectious Diseases, 1989, p 53 CRC Press, Boca Raton.).
  • the colony method is a method of taking samples and culturing them in a selective medium so that only the microorganisms to be detected can survive, and then measuring the number of colonies formed by the microorganisms. This is disadvantageous because it takes a long time to select a medium for each microorganism.
  • the DNA probe method includes a real-time PCR (polymerase chain reaction) analysis method and a nucleic acid hybridization method. After DNA microorganisms are physically destroyed, the DNA in the cell is subjected to a nucleic acid conjugation reaction. Detection method.
  • This method has the advantage that the test time is shorter than the colony method, but expensive PCR equipment is used, and when detecting a small amount of microorganisms, it is necessary to perform a separate culture step to achieve high sensitivity (Ninet, B et al. , Appl Environ Microbiol 58: 4055-4059,1992). If the incubation step is not included, dead cells can be detected, resulting in inaccurate results. In addition, when PCR is performed, false-positive is very frequent, which may increase the error range of detection and decrease the reliability of the analysis.
  • Immunoassay is an antigen-antibody binding reaction, for example, an enzyme-linked immunosorbent assay (ELISA) using an antibody that specifically reacts with the surface antigen of the microorganism to be detected is widely performed.
  • ELISA enzyme-linked immunosorbent assay
  • the present invention is to form a sensor on the membrane for filtration, to develop a sensor that combines the sensor measurement capability of the electrode and the membrane filtration characteristics.
  • An object of the present invention is to provide a sensor for measuring a biomaterial by combining an electrode measuring capability of the electrode and the separation capability of the filtration membrane by forming an electrode on the filtration membrane, a manufacturing method thereof, and a target substance detection method using the same. .
  • the present invention is a sensor electrode printed on the membrane for filtration; And it provides a sensor characterized in that to generate an electrical signal of the electrode by passing the reaction solution mixed with a sample containing the enzyme or the ligand-immobilized metal nanoparticles and the target material fixed to the filtration membrane.
  • the present invention comprises the steps of (a) printing the electrode on the membrane for filtration; And (b) passing the reaction solution in which the sample containing the enzyme or the immobilized ligand-fixed metal nanoparticles and the target substance is mixed with the filtration membrane; It provides a method of manufacturing a sensor comprising a.
  • the present invention comprises the steps of reacting by mixing a sample containing a ligand-immobilized enzyme or ligand-immobilized metal nanoparticles and the target material; (b) passing the reaction solution through a membrane for filtration on which an electrode is formed to filter out only a target substance-ligand-enzyme complex or a target substance-ligand-metal nanoparticle complex; And (c) measuring an electrical signal generated at the electrode by the target substance-ligand-enzyme complex or the target substance-ligand-metal nanoparticle complex remaining on the filtration membrane; It provides a method for detecting a target substance comprising a.
  • the present invention comprises the steps of: (a) reacting by mixing a sample containing the enzyme or the immobilized ligand-fixed metal nanoparticles and the target material; (b) a target-ligand-enzyme complex or target-ligand-metal nanoparticle that passes through the reaction solution through a filtration membrane including a receptor fixed on a filtration membrane between electrodes to specifically bind to the receptor Filtering out only the complexes; And (c) measuring an electrical signal generated at the electrode by the target substance-ligand-enzyme complex or the target substance-ligand-metal nanoparticle complex remaining on the filtration membrane; It provides a method for detecting a target substance comprising a.
  • the membrane electrode according to the present invention provides a new sensor that combines the filtration function of the membrane and the signal measurement capability of the electrode. According to the present invention, not only the target material can be measured by filtering on the membrane, but also a small amount of the target material can be detected with excellent sensitivity by using an amplified electrical signal by increasing the electrical conductivity by reducing metal ions on the membrane. It can also be useful for quantitative analysis of target substances.
  • the sample may be passed through a membrane to filter out only a target material that selectively binds to the receptor, and then may be used for electrical signal detection.
  • the signal measurement of the sensor according to the present invention can use a variety of methods, such as electrical conductivity, impedance.
  • FIG. 1 is a diagram showing a membrane electrode with an electrode spacing of 100 ⁇ m made using a silk screen printing process, and a membrane showing a membrane electrode (B, left) and a membrane carbon electrode (B, right).
  • Figure 2 is a view showing the formation process of the silver ion reduction precipitate using the enzyme in the membrane silver electrode according to the present invention.
  • FIG 3 and 4 is a diagram showing a change in current according to the presence or absence of enzymes and the reduction of silver ions in the membrane silver electrode according to the present invention.
  • FIG 5 is a view showing the formation process of the gold ion reduction precipitate using gold nanoparticles in the membrane silver electrode according to the present invention.
  • FIGS. 6 and 7 are diagrams showing a change in current according to the presence or absence of gold nanoparticles and whether the gold ion in the silver-micro electrode according to the present invention.
  • 0.1 M PB-Au Enh control without gold nanoparticles
  • AuNP-Au Enh experimental group with gold nanoparticles
  • FIG. 8 is a diagram showing the formation of gold ion reduction precipitate using the food poisoning bacteria-antibody-gold nanoparticle complex in the membrane silver electrode according to the present invention.
  • FIGS. 9 and 10 are diagrams showing the change of current according to the presence or absence of food poisoning bacteria-antibody-gold nanoparticle complex and the reduction of gold ions in the membrane electrode according to the present invention.
  • Control control group with antibody-gold nanoparticle conjugate only; 10 ⁇ 2 cell: experimental group with complex; bare-PBS: PBS addition to initial electrode; control-PBS: PBS addition after addition of gold nanoparticles; cell-PBS: food poisoning bacteria
  • -PBS addition after antibody-gold nanoparticle complex Au enh-5min: 5 min of gold reducing solution; Au enh 10min: 10 min of gold reducing solution; Au enh-PBS: PBS washing after gold reducing solution
  • FIG. 11 is a view showing a change in current according to the concentration according to the concentration of food poisoning (Staphylococcus aureus) in the membrane electrode according to the present invention.
  • Au enh-5min Gold reducing solution 5 minutes treatment
  • Au enh 10min Gold reducing solution 10 minutes treatment
  • Au enh-PBS Gold reducing solution treatment PBS wash
  • the present invention is a sensor in which the electrode is printed on the membrane for filtration; And it provides a sensor characterized in that to generate an electrical signal of the electrode by passing the reaction solution mixed with a sample containing the enzyme or the ligand-immobilized metal nanoparticles and the target material fixed to the filtration membrane.
  • the senor is printed on the membrane for filtration; And a reaction solution in which the ligand-immobilized enzyme or the metal nanoparticle in which the ligand is immobilized and the sample containing the target substance are mixed with the filtration membrane, and the reaction solution that does not bind the target substance passes through the membrane and Only the bound target-ligand-enzyme complex or target-ligand-metal nanoparticle complex remains on the membrane to generate an electrical signal of the electrode.
  • the 'filtration membrane' means a filtration membrane having pores having a diameter of 100 nm to 10 ⁇ m. Therefore, when a reaction solution obtained by reacting a ligand-immobilized enzyme or a ligand-immobilized metal nanoparticle with a sample containing a target substance is passed through a filtration membrane, the enzyme or metal nanoparticle to which the ligand is not bound is immobilized. The particles pass through the membrane, and only the target-ligand-enzyme complex or target-ligand-metal nanoparticle complex bound to the target remains on the membrane to perform a filtration function that generates a signal through an electrode formed on the membrane. have.
  • the filtration membrane may be, for example, nitrocellulose, polycarbonate, nylon, polyester, cellulose acetate, polysulfone or polyethanesulfone filtration membrane, but is not limited thereto.
  • the electrode may be prepared by a screen printing method, specifically, it may be printed by a silk screen printing process.
  • the electrode material paste platinum, gold, silver, carbon, etc.
  • the electrode material paste may be directly printed on the membrane through a screen of a predetermined pattern, and may be manufactured by repeating the drying process at a high temperature (generally 100 ° C. or more).
  • the electrode may be, for example, an interdigitated electrode (IDE).
  • IDE is an electrode in which two comb-structure electrodes face each other, and is used for detecting food poisoning bacteria by measuring the impedance method when microorganisms are coupled between the insulation of the two electrodes (Yang, L. et al. Anal. Chem , 76: 1107-1113, 2004).
  • IDE is also used as a variety of sensors, including unlabeled biosensors and gas sensors (Dharuman, V. et al. Biosens. Bioelectron., 21: 645-654, 2005, Hermans ECM, Sensor.Actuat. 5: 181- 186, 1984).
  • the interdigitated electrode may have an interval between electrodes of 10 to 1000 ⁇ m, 10 to 900 ⁇ m, 10 to 800 ⁇ m, 10 to 700 ⁇ m, 10 to 600 ⁇ m, 10 to 500 ⁇ m, 10 to 450 ⁇ m, 10 to 400 ⁇ m, 10 to 350 ⁇ m, 10 to 300 ⁇ m, 10 to 250 ⁇ m, 10 to 200 ⁇ m, or 10 to 150 ⁇ m, but are not limited thereto.
  • the 'ligand' is a substance having a specific binding force to the target substance
  • the ligand is, for example, antibodies, antigens, enzymes, peptides, proteins, DNA, RNA, peptide nucleic acids (PNA) or pressure It may be an aptamer, but is not limited thereto.
  • the type of ligand may be appropriately selected by those skilled in the art according to the type of target substance to be detected.
  • the ligand-immobilized enzyme or the ligand-immobilized metal nanoparticles and the sample containing the target substance are mixed and reacted, the ligand and the target substance and the metal nanoparticle are combined by the ligand in the mixed reaction solution.
  • a 'target-ligand-enzyme complex' or a 'target-ligand-metal nanoparticle complex' may be formed.
  • the enzyme or the ligand-immobilized metal nanoparticle to which the ligand is not bound is passed through the membrane, and the target-ligand-enzyme complex or the target-ligand-metal nanoparticle complex is bound to the target.
  • the target material can be detected by measuring this electrical signal.
  • the electrical signal to be measured is generated in proportion to the concentration of the metal nanoparticles remaining in the membrane (that is, the concentration of the target substance), thereby enabling the quantitative detection of the target substance.
  • the target material when the target material is a large microorganism, a cell or an organ of animals and plants, the target material can be easily detected even if a separate receptor is not provided on the membrane.
  • the 'target material' may be a microorganism, an antigen, a nucleic acid, a cell or an organ of an animal or plant, wherein the 'microorganism' may be, for example, a virus, a bacterium or a fungus, but is not limited thereto.
  • the 'enzyme' to which the ligand is immobilized may be, for example, peroxidase, alkaline phosphatase, galactosidase, or glucose oxidase, but is not limited thereto.
  • it may be gold, silver, copper or magnetic nanoparticles, but is not limited thereto.
  • 'nanoparticles' refers to ultrafine particles having a diameter in the range of about 1 to 100 nm.
  • nanoparticles of the size 20nm but is not limited thereto, for example, 0.5 to 100 nm, 0.5 to 90 nm, 0.5 to 80 nm, 0.5 to 70 nm, 0.5 to 70 nm, It may have a size of 0.5 to 60 nm, 0.5 to 50 nm, 0.5 to 40 nm, 0.5 to 30 nm, 0.5 to 20 nm, 1 to 20 nm.
  • a sensor can be manufactured in which the receptor is further fixed on the filtration membrane between the electrodes.
  • the senor of the present invention is further fixed to the receptor on the membrane for filtration between the electrode, the target-ligand-enzyme complex or target-ligand-metal nanoparticle complex that specifically binds to the receptor It can only be characterized by generating an electrical signal from the electrode remaining on the membrane.
  • the 'receptor' is a material having a specific binding force to the target material, characterized in that it has a different binding site than the ligand that binds the target material with enzymes or metal nanoparticles.
  • the receptor may form an electrode on the membrane and then secure it on the membrane between the electrodes. Fixing of the receptor can be carried out by methods known in the art, and both physical adsorption and chemical methods can be used. Specific immobilization method may be appropriately selected by those skilled in the art according to the type of receptor and membrane.
  • the target substance specific for the receptor binds to the receptor on the membrane.
  • the receptor-target-ligand-enzyme complex or the receptor-target-ligand-metal nanoparticle complex is immobilized on the membrane, and the target material specific for the receptor can be detected by an electrical signal.
  • the receptor may be, for example, an antibody, an antigen, an enzyme, a peptide, a protein, a DNA, an RNA, a peptide nucleic acids, or an aptamer, but is not limited thereto.
  • the target substance may be, for example, an antibody, an antigen, an enzyme, a peptide, a protein, a DNA, an RNA, a microorganism, an organ of a cell or an animal or plant, but is not limited thereto.
  • a sensor may be manufactured by reducing a metal ion on the electrode surface by adding a metal reducing solution on the filtration membrane of the sensor without the receptor or with the receptor further fixed.
  • the senor of the present invention may be characterized in that the electrode is printed on the membrane for filtration, and metal ions are reduced and deposited on the surface of the electrode.
  • the metal ions may be reduced by metal reduction solution to be deposited on the electrode surface, but the present invention is not limited thereto, and the metal ions are reduced on the electrode surface by a method appropriately selected by those skilled in the art. Can be deposited.
  • the electrical signals generated by the target-ligand-enzyme complex, target-ligand-metal nanoparticle complex, receptor-target-ligand-enzyme complex or receptor-target-ligand-metal nanoparticle complex remaining on the membrane By adding a metal reducing solution, the metal ions are reduced and deposited on the surface of the electrode to be amplified by improving electrical conductivity.
  • the 'metal reducing solution' means a solution containing a metal ion and a reducing agent capable of reducing the metal ion, and any solution having such characteristics may be used without limitation, and a specific composition may be appropriately selected by those skilled in the art.
  • the metal ion may be, for example, one or more selected from the group consisting of gold, silver, and copper, but is not limited thereto.
  • the reducing agent capable of reducing the metal ion may be selected from, for example, hydroxyl amine (NH 4 OH), ascorbic acid, glucose, and mixtures thereof, but is not limited thereto.
  • the electrical conductivity of the electrode is improved, thereby amplifying the electrical signal.
  • the reduction reaction of these metal ions is greatly promoted by enzymes or metal nanoparticles to which the ligands added on the membrane are immobilized, and the electrical conductivity is remarkably improved by the enzyme or metal nanoparticles. Very good sensors can be manufactured.
  • the present invention comprises the steps of (a) printing the electrode on the membrane for filtration; And (b) passing the reaction solution in which the sample containing the enzyme or the immobilized ligand-fixed metal nanoparticles and the target substance is mixed with the filtration membrane; It provides a method of manufacturing a sensor comprising a.
  • step (a) may further comprise the step of fixing the receptor further on the membrane for filtration between the electrodes.
  • the method may further include the step of reducing the metal ion to the electrode surface by adding a metal reducing solution on the filtration membrane of the sensor without the receptor or with the receptor further fixed.
  • the electrode, ligand, enzyme, nanoparticle, target substance, receptor, metal reducing solution and the like are the same as the sensor.
  • the present invention comprises the steps of reacting by mixing a sample containing a ligand-immobilized enzyme or ligand-immobilized metal nanoparticles and the target material; (b) passing the reaction solution through a membrane for filtration on which an electrode is formed to filter out only a target substance-ligand-enzyme complex or a target substance-ligand-metal nanoparticle complex; And (c) measuring an electrical signal generated at the electrode by the target substance-ligand-enzyme complex or the target substance-ligand-metal nanoparticle complex remaining on the filtration membrane; It provides a method for detecting a target substance comprising a.
  • the present invention comprises the steps of: (a) reacting by mixing a sample containing the enzyme or the immobilized ligand-fixed metal nanoparticles and the target material; (b) a target-ligand-enzyme complex or target-ligand-metal nanoparticle that passes through the reaction solution through a filtration membrane including a receptor fixed on a filtration membrane between electrodes to specifically bind to the receptor Filtering out only the complexes; And (c) measuring an electrical signal generated at the electrode by the target substance-ligand-enzyme complex or the target substance-ligand-metal nanoparticle complex remaining on the filtration membrane; It provides a method for detecting a target substance comprising a.
  • the method may further include the step of reducing and depositing metal ions on the electrode surface by adding a metal reducing solution on the filtration membrane before measuring the electrical signal.
  • the enzyme or a metal may be amplified by an electrically measurable signal and measured.
  • the measurement of the signal may be an electrical signal measured through an electrode on the membrane, for example, but may be by measuring the electrical conductivity or impedance, but is not limited thereto.
  • the object detection method of the present invention can be performed using the above-described sensor, it can be seen to include both the configuration and features of the above-described sensor.
  • An interlocked electrode of 100 ⁇ m spacing was fabricated using silk screen printing technology using silver paste on an MMM (asymetric super-micron membrane) membrane with 0.45 ⁇ m pores.
  • FIG. 2 is a diagram illustrating a process of fixing peroxidase enzyme (HRP) to a membrane silver electrode prepared by the above-described method and reducing silver ions in the silver reducing solution prepared by the above-described method on the electrode by the peroxidase enzyme. .
  • HRP peroxidase enzyme
  • 3 and 4 is a membrane prepared by the above-described method to fix the peroxidase enzyme to the electrode, the silver is reduced by using the enzyme and the silver reducing solution prepared by the method described above, after applying a DC voltage to the current Shows the result of measuring the change in.
  • STA / HRP-AgGSH peroxidase enzyme
  • PB-AgGSH control group without peroxidase enzyme
  • An interlocked electrode of 100 ⁇ m spacing was fabricated using silk screen printing technology using silver paste on an MMM (asymetric super-micron membrane) membrane with 0.45 ⁇ m pores.
  • FIG. 5 is a diagram illustrating a process of fixing gold nanoparticles to a membrane silver electrode prepared by the above-described method and reducing the gold reducing solution prepared by the above-described method on the electrode by gold nanoparticles.
  • 6, 7 is a membrane prepared by the above-described method to fix the gold nanoparticles to the micro-electrode, and to reduce the gold by using a gold reducing solution prepared by the above-described method, applying a DC voltage thereto to the current Show the results of measuring changes.
  • the electrical conductivity was reduced by treating gold reducing solution (5min, 10min). It can be seen that is significantly improved.
  • Example 3 Membrane Ion Reduction After Filtering Food Poisoning-Antibody-Gold Nanoparticle Complex to Electrode
  • BB International 20 nm gold nanoparticle solution
  • borate buffer 0.1 M, pH 8.5
  • a selective antibody Abcam, ab20002
  • BSA bovine serum albumin
  • Example 3-2 Staphylococcus aureus culture
  • the membrane prepared by the method described above was filtered through a micro-electrode. Gold ions were reduced between the electrodes by reacting 10 mM citrate buffer (pH 3.0) (gold reducing solution) containing 1 mM hydroxyl amine and 10 mM HAuCl 4. The current was measured by applying a DC voltage thereto.
  • Figure 8 is a membrane prepared by the above-described method is fixed to the food poisoning bacteria-gold nanoparticle complex on the micro-electrode, the gold reducing solution prepared by the above-described method is shown the process of reduction on the electrode by the gold nanoparticles.
  • 9 and 10 is a membrane prepared by the above-described method is fixed to the food poisoning bacteria-gold nanoparticle complexes on the micro-electrode, after reducing the gold using the gold reducing solution prepared by the above-described method, the DC voltage It shows the result of measuring the change of the walking current.
  • FIG. 11 shows current values at 0.1 V according to Staphylococcus aureus concentrations from the experiments of FIGS. 9 and 10. It is shown that the current value increases with the concentration of bacteria, and it is possible to measure the bacteria concentration of about 102 cfu.
  • the sample may be passed through a membrane to filter out only a target material that selectively binds to the receptor, and then may be used for electrical signal detection.
  • the signal measurement of the sensor according to the present invention can use a variety of methods, such as electrical conductivity, impedance.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Pathology (AREA)
  • Urology & Nephrology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Hematology (AREA)
  • General Physics & Mathematics (AREA)
  • Biomedical Technology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Nanotechnology (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • Electrochemistry (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)

Abstract

La présente invention concerne un procédé de détection de biomolécule au cours duquel une électrode est formée sur une membrane de filtration par impression sérigraphique pour combiner les caractéristiques de filtration de la membrane et la capacité de mesure de signaux de l'électrode. Une électrode à membrane selon la présente invention fournit un nouveau capteur dans la lequel la fonction de filtration de la membrane et la capacité de mesure de signaux de l'électrode sont combinées. Selon la présente invention, un matériau cible est filtré par la membrane et mesuré, et un ion métallique est réduit sur la membrane, ce qui permet d'augmenter la conductibilité électrique, ce qui a pour conséquence que la très petite quantité de matériau cible peut être détectée avec une sensibilité supérieure au moyen des signaux électriques amplifiés, et une analyse quantitative du matériau cible peut être effectuée de manière avantageuse au moyen des signaux électriques amplifiés. En outre, un matériau récepteur peut être fixé sur l'électrode précédemment mentionnée, l'échantillon ainsi formé passant à travers la membrane pour obtenir uniquement le matériau cible se liant de manière sélective au matériau récepteur, et le matériau cible ainsi obtenu peut être utilisé pour la détection de signaux électriques. La mesure des signaux du capteur selon la présente invention peut utiliser divers éléments tels que la conductivité électrique et l'impédance.
PCT/KR2011/009746 2010-12-16 2011-12-16 Impression au moyen d'une électrode à phase membranaire et détection de biomolécule utilisant ladite impression Ceased WO2012081940A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US13/994,348 US9465003B2 (en) 2010-12-16 2011-12-16 Membrane phase electrode using printing and bio-molecule detection using same
US15/269,779 US20170010234A1 (en) 2010-12-16 2016-09-19 Membrane phase electrode using printing and bio-molecule detection using same

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2010-0128965 2010-12-16
KR20100128965 2010-12-16
KR1020110136334A KR101335246B1 (ko) 2010-12-16 2011-12-16 프린팅을 이용한 멤브레인 상 전극 및 이를 활용한 생체물질 검출
KR10-2011-0136334 2011-12-16

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US13/994,348 A-371-Of-International US9465003B2 (en) 2010-12-16 2011-12-16 Membrane phase electrode using printing and bio-molecule detection using same
US15/269,779 Division US20170010234A1 (en) 2010-12-16 2016-09-19 Membrane phase electrode using printing and bio-molecule detection using same

Publications (2)

Publication Number Publication Date
WO2012081940A2 true WO2012081940A2 (fr) 2012-06-21
WO2012081940A3 WO2012081940A3 (fr) 2012-10-04

Family

ID=46245248

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2011/009746 Ceased WO2012081940A2 (fr) 2010-12-16 2011-12-16 Impression au moyen d'une électrode à phase membranaire et détection de biomolécule utilisant ladite impression

Country Status (1)

Country Link
WO (1) WO2012081940A2 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105372423A (zh) * 2015-11-22 2016-03-02 长沙市食品质量安全监督检测中心 基于微间距阵列电极的免疫定量传感器检测金黄色葡萄球菌的试剂盒及方法

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1043588A4 (fr) * 1997-12-25 2001-08-22 Mochida Pharm Co Ltd Analyseur pour echantillons liquides
US6576460B1 (en) * 1999-10-28 2003-06-10 Cornell Research Foundation, Inc. Filtration-detection device and method of use
KR100427599B1 (ko) * 2001-05-30 2004-04-27 주식회사 아이센스 대면형 다공성 전극을 포함하는 자가 시료채취 흐름계형바이오 센서
EP1512010B1 (fr) * 2002-05-31 2008-01-02 Cornell Research Foundation, Inc. Biocapteur universel et procedes d'utilisation
CA2493101C (fr) * 2002-07-20 2012-05-08 Acea Biosciences, Inc. Appareils a base d'impedance et procedes d'analyse de cellules et de particules

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105372423A (zh) * 2015-11-22 2016-03-02 长沙市食品质量安全监督检测中心 基于微间距阵列电极的免疫定量传感器检测金黄色葡萄球菌的试剂盒及方法

Also Published As

Publication number Publication date
WO2012081940A3 (fr) 2012-10-04

Similar Documents

Publication Publication Date Title
KR101335246B1 (ko) 프린팅을 이용한 멤브레인 상 전극 및 이를 활용한 생체물질 검출
Eissa et al. Ultrasensitive peptide-based multiplexed electrochemical biosensor for the simultaneous detection of Listeria monocytogenes and Staphylococcus aureus
Dai et al. Electrochemical determination of Salmonella typhimurium by using aptamer-loaded gold nanoparticles and a composite prepared from a metal-organic framework (type UiO-67) and graphene
Rishpon et al. An amperometric enzyme-channeling immunosensor
Lin et al. Disposable amperometric immunosensing strips fabricated by Au nanoparticles-modified screen-printed carbon electrodes for the detection of foodborne pathogen Escherichia coli O157: H7
Pérez et al. Immunomagnetic separation with mediated flow injection analysis amperometric detection of viable Escherichia coli O157
Gattani et al. Recent progress in electrochemical biosensors as point of care diagnostics in livestock health
Paniel et al. Colorimetric and electrochemical genosensors for the detection of Escherichia coli DNA without amplification in seawater
Li et al. A dual-signal amplification strategy for kanamycin based on ordered mesoporous carbon-chitosan/gold nanoparticles-streptavidin and ferrocene labelled DNA
CN104764790B (zh) 基于核酸适配体检测链霉素的生物传感器及其制备方法
US20090061524A1 (en) Enzyme-Channeling Based Electrochemical Biosensors
WO2011149233A2 (fr) Composite de nanoparticules magnétiques/nanoparticules de platine/carbone poreux et procédé pour sa préparation
Kheiri et al. A novel amperometric immunosensor based on acetone-extracted propolis for the detection of the HIV-1 p24 antigen
CN108760853A (zh) 一种检测牛奶中卡那霉素残留的适配体传感器的制备方法
CN101532980B (zh) 检测志贺氏菌的酶免疫传感器及其制备方法和运用
Hu et al. A lab-on-paper biosensor using a two-step enzymatic amplification strategy for ultrasensitive detection of active BoNT/A in complex matrices
CN115356480A (zh) 一种糖基化磁性类细胞识别受体的制备方法
Jiang et al. An electrochemical strategy with molecular beacon and hemin/G-quadruplex for the detection of Clostridium perfringens DNA on screen-printed electrodes
La Belle et al. Label‐free and ultra‐low level detection of Salmonella enterica serovar Typhimurium using electrochemical impedance spectroscopy
US20230296551A1 (en) Method for detecting and quantifying analytes in a microfluidic device
US7241626B2 (en) Isolation and confirmation of analytes from test devices
Xu et al. Simultaneous detection of Staphylococcus aureus and E. coli O157: H7 using a self-calibrated potentiometric sensors array based on peptide recognition
Vetcha et al. Detection of hantavirus infection in hemolyzed mouse blood using alkaline phosphatase conjugate
Yang et al. Immunoelectrochemical Assay in Combination with Homogeneous Enzyme‐Labeled Antibody Conjugation for Rapid Detection of Salmonella
US10768177B2 (en) Bacteriophage-based electrochemical biosensor

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11849253

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 13994348

Country of ref document: US

122 Ep: pct application non-entry in european phase

Ref document number: 11849253

Country of ref document: EP

Kind code of ref document: A2