CN112611793A - Preparation method of paper-based dual-mode amyloid biosensor - Google Patents

Preparation method of paper-based dual-mode amyloid biosensor Download PDF

Info

Publication number
CN112611793A
CN112611793A CN202110000827.2A CN202110000827A CN112611793A CN 112611793 A CN112611793 A CN 112611793A CN 202110000827 A CN202110000827 A CN 202110000827A CN 112611793 A CN112611793 A CN 112611793A
Authority
CN
China
Prior art keywords
area
paper
ceo
cuco
electrode
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.)
Withdrawn
Application number
CN202110000827.2A
Other languages
Chinese (zh)
Inventor
崔康
周晨曦
李旭
于京华
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.)
University of Jinan
Original Assignee
University of Jinan
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 University of Jinan filed Critical University of Jinan
Priority to CN202110000827.2A priority Critical patent/CN112611793A/en
Publication of CN112611793A publication Critical patent/CN112611793A/en
Withdrawn legal-status Critical Current

Links

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
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • 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
    • G01N27/301—Reference 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
    • G01N27/38—Cleaning of electrodes

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Immunology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Electrochemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Nanotechnology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Plasma & Fusion (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

The invention provides a preparation method of a high-performance paper-based dual-mode amyloid biosensor, and belongs to the technical field of biosensor preparation. In the course of the invention, a novel paper analysis device (oPAD) was prepared, which modifies Cu/Co-doped CeO with the aid of Pd2（CuCo‑CeO2-Pd) nanospheres, and the functional oPAD is constructed by combining the electrocatalytic enhancement effect of the nanostructure synergy of the-Pd) nanospheres, so that an electrochemical and chromogenic signal reading system in the oPAD can be realized, and the functional oPAD nanospheres are used for detecting amyloid protein with high sensitivity. Introduction of CuCo-CeO2The purpose of the Pd nanospheres is to design them as enhanced "signal transducer layers", furthermore, in order to realize the proposed oPADThe method of growing gold nanoparticles on cellulose fibers in situ is adopted to improve the 'identification layer', and finally, the dual-mode signal reading can be realized by simply converting the spatial configuration of the paper-based biosensor, and electrochemical measurement and colorimetric detection are sequentially performed on a compatible paper-based working electrode.

Description

Preparation method of paper-based dual-mode amyloid biosensor
Technical Field
The invention relates to a preparation method of a paper-based dual-mode amyloid biosensor, and belongs to the technical field of biosensor preparation.
Background
Aggregation of amyloid proteins with amino acid residues can cause neuronal damage and further lead to the development of associated symptoms. New protocols and strategies for the detection of amyloid in body fluids are developed and ultimately it is crucial to implement methods for amyloid detection in elderly people in ultra-sensitive, cost-effective portable devices.
In order to meet the urgent need of amyloid analysis strategies, several techniques including fluorescence technology, surface enhanced raman spectroscopy, enzyme-linked immunosorbent assay have been developed. Although advances have been made, there still exist many technical challenges in the development of analytical equipment for early diagnosis, such as complex instrument requirements, complicated procedures, low sensitivity of quantitative measurement, etc., which all increase the difficulty of routine and rapid detection.
The paper-based analysis device has the advantages of paper cellulose, such as low cost, abundant reserves, easy functionalization, green biodegradation and the like, so that the paper-based analysis device has great attention in nursing and diagnosis. In view of its attractive properties, paper analysis devices have been integrated with various technologies, such as paper-based fluorescence, paper-based electrochemiluminescence, and paper-based colorimetry, and the like, and have been successfully used in environmental monitoring, energy storage applications, and the like. In particular, the combination of electrochemical techniques with paper-based analytical devices makes them attractive compared to other methods due to their high sensitivity, selectivity and simplicity, and is a potential analytical method for the detection of amyloid.
Disclosure of Invention
Aiming at the problems existing at present, the technical problem to be solved by the invention is to provide a preparation method of a paper-based dual-mode amyloid biosensor, which is reasonable in design, low in cost, simple and convenient to operate and high in sensitivity, and is characterized by comprising the following steps:
(1) preparation of soluble amyloid by first obtaining amyloid by dissolving lyophilized amyloid peptide in 1,1,1,3,3,3, 3-hexafluoro-2-propanol and sonicating for 10 min, then re-dissolving the obtained product in 20 mM sodium hydroxide solution to obtain a concentration of 2-3 mM, preparing a sample by diluting to a final concentration of 50-60 μ M with 10-20 mM phosphate buffer, pH 2.0, and further diluting to a desired concentration with 2-3 mM phosphate buffer, pH 7.2-7.5, for further experiments at 25 ℃;
(2) Ab2/CuCo-CeO2preparation of Pd bioconjugates: firstly, 1.0-1.5 mg of CuCo-CeO2the-Pd nanospheres were dispersed in 200-300. mu.L of pH 7.4 phosphate buffer, then transferred to 200-300. mu.L of 1.0. mu.g/mL solution of Ab2 and incubated at 4 ℃ for 24 h, and then the prepared Ab2/CuCo-CeO was collected by centrifugation2Pd bioconjugates and further washing with phosphate buffer pH 7.4, Ab2/CuCo-CeO once obtained in order to prevent surface non-specific active sites2Pd bioconjugates, 100-150. mu.L of 1.0% -1.5% bovine serum albumin was added to the obtained product and shaken at 4 ℃ for 3 hours, then excess bovine serum albumin was washed away with a phosphate buffer solution of pH 7.4, and the prepared bioconjugates were stored in a phosphate buffer solution of pH 7.4 in the presence of 4 ℃ for the next step of the experiment;
(3) designing and assembling the paper-based biosensor: designing a hydrophobic wax printing pattern on a computer by using Adobe Illustrator CS6 software, printing the hydrophobic wax printing pattern on cut A4-sized filter paper in batch by using a wax-spraying printer, printing a working electrode, a counter electrode and a reference electrode on corresponding unprinted circular area paper fibers on the basis of a screen printing technology, wherein the pattern is as shown in the attached drawing 1, and in order to finally complete the assembly of the three-dimensional structure of the paper-based biosensor, cutting the printed paper into different single labels by using a razor, and then folding each label along the connecting space between the two labels;
(4) functionalization of the detection area: firstly, growing gold nanoparticles by a seed solution growth method, wherein the specific growth steps are as follows: firstly pouring 80-100 mL of secondary water into a three-neck flask, heating to 90 ℃, adding 500-1000 mu L of chloroauric acid solution with the mass fraction of 1% -2%, continuously heating to 96 ℃ and keeping for 1 min, finally adding 2.0-3.0 mL of sodium citrate with the mass fraction of 1%, continuously heating for 15 min, naturally cooling to obtain gold seed solution, dropwise adding 50-100 mu L of gold seed solution into a working area, standing, airing and dryingRepeated three times, washing with secondary water for 3 times, then dropping 25 μ L of 20-25 μ g/mL Ab1 on the prepared gold-paper electrode, incubating at 4 deg.C for 12 h, washing with pH 7.4 phosphate buffer solution for 3 times, blocking the resulting electrode with 25 μ L of 2.0% -3.0% bovine serum albumin for 2 h at room temperature, and preparing Ab2/CuCo-CeO by antigen-antibody specific interaction for electrochemical signal amplification2CuCo-CeO of-Pd bioconjugates2the-Pd probe is introduced into the system, in which CuCo-CeO2The unique catalytic properties of Pd help to effectively increase the sensitivity of target detection during bioanalysis, and then the gold-paper electrode in the detection zone was smoothly washed with phosphate buffer pH 7.0 to remove unbound Ab2/CuCo-CeO2-a Pd bioconjugate;
(5) pretreatment of color reaction: adding 10 mu L of phosphate buffer solution with the pH value of 7.4 and the concentration of 0.1-0.5M into the circular hydrophilic area, then dropwise adding 200 mu L of hydrogen peroxide solution with the concentration of 5-8 mM into the circular hydrophilic area, and then pre-burying 20 mu L of 3,3',5,5' -tetramethylbenzidine color developing solution with the concentration of 20-30 mM in the color developing area;
(6) detection of electrochemical signals: firstly, overlapping a detection area, a reference electrode area, a counter electrode area and a shunt area by folding, arranging a cleaning area below a color development area, after finishing the layer-by-layer modification of the electrodes, cleaning a working area to ensure that the detection area and an electrode label are overlapped, fixing by a clamp, connecting with an electrochemical workstation, wherein 60 mu L of the electrochemical workstation contains 5.0-6.0 mM [ Fe (CN)6]3-/4-Dropping phosphate buffer solution with pH of 7.4 and concentration of 0.1M into the detection area, and measuring and recording;
(7) measurement of color development Signal: after the electric signal measurement process is finished, the cleaning label is drawn off, the style is shown in figure 3, hydrogen peroxide and the signal probe flow through the shunt area to flow to the color development area and the contrast area respectively and react with 3,3',5,5' -tetramethyl benzidine color development liquid pre-buried in the color development area, so as to realize visual detection.
The design of the paper chip in the step (3) is characterized in that: the clean region is hydrophobic region, the size is 12 mm x 12 mm, the regional diameter of working electrode is 10 mm, the modification region diameter is 8 mm, reference electrode and counter electrode printing region diameter are 10 mm, the design has constructed a reposition of redundant personnel region, the diameter is 10 mm, can realize the reposition of redundant personnel of reaction liquid, make it flow respectively to the contrast that color development region and contrast region carry out color development reaction, color development region and contrast region diameter are 10 mm, the size of waste liquid pond is 18 mm x 8 mm in the washing label, can realize electrode modification in-process and wash many times.
The invention has the beneficial effects that:
(1) a preparation method of a paper-based dual-mode amyloid biosensor reduces experiment cost.
(2) The introduction of the signaling probe greatly improves the selectivity and sensitivity of the detection.
(3) The functionalization of the paper-based gold nanoparticles can effectively increase the specific surface area of the paper-based gold nanoparticles and improve the detection sensitivity.
(4) The paper-based sensor is flexible and flexible, convenient to carry, capable of being cut, bent, folded and plastic, simple in post-processing and free of environmental pollution.
(5) Compared with the traditional glassy carbon electrode and glass electrode, the paper substrate has the advantages of rich raw materials, light weight, low price, easy folding and degradability.
(6) The paper-based biosensor realizes the self-cleaning process of the electrode, simplifies the operation steps and saves the electrode modification time.
Description of the drawings:
the invention is described in further detail below with reference to the figures and specific embodiments:
FIG. 1 shows a working electrode, a reference electrode, a counter electrode, and carbon lines printed on a hydrophobic wax pattern.
FIG. 2 is a schematic diagram of the folding of an electrochemical-mode detection target of a paper-based device.
FIG. 3 is a schematic view showing the folding of the target object detected by the color development mode of the paper-based device.
Detailed Description
The preparation method of the paper-based dual-mode amyloid biosensor is reasonable in design, low in cost, simple and convenient to operate and high in sensitivity, and is characterized by comprising the following steps of:
(1) preparation of soluble amyloid by first obtaining amyloid by dissolving lyophilized amyloid peptide in 1,1,1,3,3,3, 3-hexafluoro-2-propanol and sonicating for 10 min, then redissolving the obtained product in 20 mM sodium hydroxide solution to a concentration of 2 mM, preparing a sample by diluting to a final concentration of 50 μ M with 10 mM, pH 2.0 phosphate buffer, and further diluting to a desired concentration with 2 mM, pH 7.2 phosphate buffer for further experiments at 25 ℃;
(2) Ab2/CuCo-CeO2preparation of Pd bioconjugates: firstly, 1.0 mg of CuCo-CeO2Pd nanospheres dispersed in 250. mu.L of pH 7.4 phosphate buffer, then transferred to 250. mu.L of 1.0. mu.g/mL solution of Ab2 and incubated at 4 ℃ for 24 h, and the prepared Ab2/CuCo-CeO was collected by centrifugation2Pd bioconjugates and further washing with phosphate buffer pH 7.4, Ab2/CuCo-CeO once obtained in order to prevent surface non-specific active sites2Pd bioconjugates, 150. mu.L of 1.0% bovine serum albumin was added to the obtained product and shaken at 4 ℃ for 3 h, then excess bovine serum albumin was washed out with phosphate buffer pH 7.4, and the prepared bioconjugates were stored in phosphate buffer pH 7.4 for further experiments at 4 ℃;
(3) designing and assembling the paper-based biosensor: designing a hydrophobic wax printing pattern on a computer by using Adobe Illustrator CS6 software, printing the hydrophobic wax printing pattern on cut A4-sized filter paper in batch by using a wax-spraying printer, printing a working electrode, a counter electrode and a reference electrode on corresponding unprinted circular area paper fibers on the basis of a screen printing technology, wherein the pattern is as shown in the attached drawing 1, and in order to finally complete the assembly of the three-dimensional structure of the paper-based biosensor, cutting the printed paper into different single labels by using a razor, and then folding each label along the connecting space between the two labels;
(4) functionalization of the detection area: firstly, growing gold nanoparticles by a seed solution growth method, and concretely growing the gold nanoparticlesComprises the following steps: pouring 80 mL of secondary water into a three-neck flask, heating to 90 ℃, adding 800 μ L of chloroauric acid solution with the mass fraction of 1%, continuously heating to 96 ℃ for 1 min, finally adding 2.8 mL of sodium citrate with the mass fraction of 1%, continuously heating for 15 min, naturally cooling to obtain gold seed solution, dropwise adding 60 μ L of the obtained gold seed solution into a working area, standing, airing, repeating for three times, washing with the secondary water for 3 times, then, 25. mu.L of 24. mu.g/mL Ab1 was dropped onto the prepared gold-paper electrode, and incubated at 4 ℃ for 12 h, washed 3 times with phosphate buffer pH 7.4, the resulting electrode was blocked with 25. mu.L of 2.0% bovine serum albumin for 2 h at room temperature, and in order to achieve electrochemical signal amplification, it will be prepared as Ab2/CuCo-CeO in advance by antigen-antibody specific interaction.2CuCo-CeO of-Pd bioconjugates2the-Pd probe is introduced into the system, in which CuCo-CeO2The unique catalytic properties of Pd help to effectively increase the sensitivity of target detection during bioanalysis, and then the gold-paper electrode in the detection zone was smoothly washed with phosphate buffer pH 7.0 to remove unbound Ab2/CuCo-CeO2-a Pd bioconjugate;
(5) pretreatment of color reaction: adding 10 mu L of phosphate buffer solution with the pH value of 7.4 and the concentration of 0.1M into the circular hydrophilic area, then dropwise adding 200 mu L of hydrogen peroxide solution with the concentration of 5 mM into the circular hydrophilic area, and then pre-burying 20 mu L of 3,3',5,5' -tetramethylbenzidine color developing solution with the concentration of 20 mM in the color developing area;
(6) detection of electrochemical signals: firstly, overlapping a detection area, a reference electrode area, a counter electrode area and a shunt area by folding, arranging a cleaning area below a color development area, after finishing the layer-by-layer modification of the electrodes, cleaning a working area to ensure that the detection area and an electrode label are overlapped, fixing by a clamp, connecting with an electrochemical workstation, and connecting 60 mu L of the electrode with 5.0 mM of Fe (CN)6]3-/4-Dropping phosphate buffer solution with pH of 7.4 and concentration of 0.1M into the detection area, and measuring and recording;
(7) measurement of color development Signal: after the electric signal measurement process is finished, the cleaning label is drawn off, the style is shown in figure 3, hydrogen peroxide and the signal probe flow through the shunt area to flow to the color development area and the contrast area respectively and react with 3,3',5,5' -tetramethyl benzidine color development liquid pre-buried in the color development area, so as to realize visual detection.

Claims (2)

1. A preparation method of a high-performance paper-based dual-mode amyloid biosensor is characterized by comprising the following steps:
(1) preparation of soluble amyloid protein by first dissolving lyophilized amyloid peptide in 1,1,1,3,3,3, 3-hexafluoro-2-propanol and ultrasonic treatment for 10 min to obtain amyloid protein, then the obtained product is dissolved again in 20 mM sodium hydroxide solution, obtained concentration of 2 mM, by using 10 mM, pH 2.0 phosphate buffer solution to the final concentration of 50 u M, further with 2 mM, pH 7.2 phosphate buffer solution diluted to the desired concentration of preparation sample, in order to carry out further experiments at 25 ℃;
(2) Ab2/CuCo-CeO2preparation of Pd bioconjugates: firstly, 1.0 mg of CuCo-CeO2Pd nanospheres dispersed in 250. mu.L of pH 7.4 PBS solution, then transferred to 250. mu.L of 1.0. mu.g/mL solution of Ab2 and incubated at 4 ℃ for 24 h, and the prepared Ab2/CuCo-CeO was collected by centrifugation2Pd bioconjugate and further washed with 2 mM pH 7.4, Ab2/CuCo-CeO once obtained in order to prevent surface non-specific active sites2Pd bioconjugates, 150. mu.L of 1.0% bovine serum albumin was added to the obtained product and shaken at 4 ℃ for 3 h, then excess bovine serum albumin was washed out with phosphate buffer pH 7.4, and the prepared bioconjugates were stored in phosphate buffer pH 7.4 for further experiments at 4 ℃;
(3) designing and assembling the paper-based biosensor: designing a hydrophobic wax printing pattern on a computer by using Adobe Illustrator CS6 software, printing the hydrophobic wax printing pattern on cut A4-sized filter paper in batch by using a wax-spraying printer, printing a working electrode, a counter electrode and a reference electrode on corresponding unprinted circular area paper fibers on the basis of a screen printing technology, cutting the printed paper into different single labels by using a razor in order to finally finish the assembly of the three-dimensional structure of the paper-based biosensor, and folding each label along a connecting space between the two labels;
(4) functionalization of the detection area: firstly, growing gold nanoparticles by a seed solution growth method, wherein the specific growth steps are as follows: pouring 80 mL of secondary water into a three-neck flask, heating to 90 ℃, adding 800 μ L of chloroauric acid solution with the mass fraction of 1%, continuously heating to 96 ℃ for 1 min, finally adding 2.8 mL of sodium citrate with the mass fraction of 1%, continuously heating for 15 min, naturally cooling to obtain gold seed solution, dropwise adding 60 μ L of the obtained gold seed solution into a working area, standing, airing, repeating for three times, washing with the secondary water for 3 times, then, 25. mu.L of 24. mu.g/mL Ab1 was dropped onto the prepared gold-paper electrode, and incubated at 4 ℃ for 12 h, washed 3 times with phosphate buffer pH 7.4, the resulting electrode was blocked with 25. mu.L of 2.0% bovine serum albumin for 2 h at room temperature, and in order to achieve electrochemical signal amplification, it will be prepared as Ab2/CuCo-CeO in advance by antigen-antibody specific interaction.2CuCo-CeO of-Pd bioconjugates2the-Pd probe is introduced into the system, in which CuCo-CeO2The unique catalytic properties of Pd help to effectively increase the sensitivity of target detection during bioanalysis, and then the gold-paper electrode in the detection zone was smoothly washed with phosphate buffer pH 7.0 to remove unbound Ab2/CuCo-CeO2-a Pd bioconjugate;
(5) pretreatment of color reaction: adding 10 mu L of phosphate buffer solution with the pH value of 7.4 and the concentration of 0.1M into the circular hydrophilic area, then dropwise adding 200 mu L of hydrogen peroxide solution with the concentration of 5 mM into the circular hydrophilic area, and then pre-burying 20 mu L of 3,3',5,5' -tetramethylbenzidine color developing solution with the concentration of 20 mM in the color developing area;
(6) detection of electrochemical signals: firstly, overlapping a detection area, a reference electrode area, a counter electrode area and a shunt area by folding, arranging a cleaning area below a color development area, cleaning a working area after finishing layer-by-layer modification of the electrodes, overlapping the detection area and an electrode label, fixing by a clamp, connecting with an electrochemical workstation, and adding 60 mu L of a solution containing 5.0 mM of [ Fe (CN)6]3-/4-Dropping phosphate buffer solution with pH of 7.4 and concentration of 0.1M into the detection area, and measuring and recording;
(7) measurement of color development Signal: after the electric signal measurement process is finished, the cleaning label is drawn out, the hydrogen peroxide and the signal probe flow through the shunt area to the color development area and the contrast area respectively and react with the 3,3',5,5' -tetramethyl benzidine color development liquid pre-buried in the color development area, so that visual detection is realized.
2. The preparation method of the high-performance paper-based bimodal amyloid biosensor as claimed in claim 1, wherein the clean area in the assembled paper-based biosensor designed in step (3) is a hydrophobic area with a size of 12 mm x 12 mm, the diameter of the working electrode area is 10 mm, the diameter of the modification area is 8 mm, the diameters of the reference electrode and counter electrode printing areas are 10 mm, a shunting area with a diameter of 10 mm is designed and constructed, so that shunting of the reaction liquid can be realized, the reaction liquid can flow to the color development area and the contrast area respectively for contrast of color development reaction, the diameters of the color development area and the contrast area are 10 mm, the size of a waste liquid pool in the washing label is 18 mm x 8 mm, and multiple washing in the electrode modification process can be realized.
CN202110000827.2A 2021-01-04 2021-01-04 Preparation method of paper-based dual-mode amyloid biosensor Withdrawn CN112611793A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110000827.2A CN112611793A (en) 2021-01-04 2021-01-04 Preparation method of paper-based dual-mode amyloid biosensor

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110000827.2A CN112611793A (en) 2021-01-04 2021-01-04 Preparation method of paper-based dual-mode amyloid biosensor

Publications (1)

Publication Number Publication Date
CN112611793A true CN112611793A (en) 2021-04-06

Family

ID=75253263

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110000827.2A Withdrawn CN112611793A (en) 2021-01-04 2021-01-04 Preparation method of paper-based dual-mode amyloid biosensor

Country Status (1)

Country Link
CN (1) CN112611793A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114166918A (en) * 2021-12-08 2022-03-11 南开大学 Method for detecting total content of malachite green and metabolic products thereof in aquatic products by one-step method
WO2023020434A1 (en) * 2021-08-17 2023-02-23 南京岚煜生物科技有限公司 Electrochemical detection method based on screen-printed electrode

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023020434A1 (en) * 2021-08-17 2023-02-23 南京岚煜生物科技有限公司 Electrochemical detection method based on screen-printed electrode
CN114166918A (en) * 2021-12-08 2022-03-11 南开大学 Method for detecting total content of malachite green and metabolic products thereof in aquatic products by one-step method
CN114166918B (en) * 2021-12-08 2024-03-22 南开大学 Method for detecting malachite green and total metabolite thereof in aquatic product by one-step method

Similar Documents

Publication Publication Date Title
Jiao et al. 3D vertical-flow paper-based device for simultaneous detection of multiple cancer biomarkers by fluorescent immunoassay
CN112611793A (en) Preparation method of paper-based dual-mode amyloid biosensor
CN103760209B (en) Multiparameter paper chip electrochemical immunosensor
CN100520366C (en) SERS biological probe and method for making same
Carneiro et al. Biosensors on the road to early diagnostic and surveillance of Alzheimer's disease
CN111505281A (en) Sensitization type immunochromatography kit for sensitively detecting novel coronavirus antibody
CN106596969A (en) Production method, product, detection method and application of electrochemiluminescence immunosensor
CN102854324B (en) Method for rapid nondestructive detection of liver tumor marker and test paper strip adopted by the method
CN104977277B (en) It is a kind of to detect the nano vesicle of intracellular wild type and variation p53 albumen simultaneously
CN109975375A (en) A kind of zearalenone detection method based on signal enhancing type polymer functionalization red schorl phase titanium dioxide mesomorphic
US20050176067A1 (en) Biosensor for determining an allergen with operating procedures
CN102087285B (en) Immunochromatographic test strip for rapidly detecting acute pancreatitis and preparation method thereof
CN204925133U (en) Kidney dysfunction fluorescent quantitation detector
CN110501405A (en) A kind of method of integrated paper base dual-mode biological sensor detection miRNA-155
CN103995112B (en) For the test strips and preparation method thereof of specificity animal wool dandruff allergens IgE examination
Ling et al. A label-free visual immunoassay on solid support with silver nanoparticles as plasmon resonance scattering indicator
Almajidi et al. Recent applications of microfluidic immunosensors
CN103983789B (en) For the test strips and preparation method thereof of specificity dirt mite class anaphylactogen IgE examination
CN103995116B (en) For the test strips and preparation method thereof of specificity cephalosporins anaphylactogen IgE examination
CN212586389U (en) Time-resolved fluorescence immunochromatography kit for detecting novel coronavirus
Vestergaard et al. The study of Alzheimer’s disease biomarkers: Current role and future prospects of nanosensor technology
Chen et al. Accurate and portable tumor exosomes detection based on manganese dioxide and aptamer-functionalized fluorescent microspheres mediated dual-mode lateral flow assay
CN103995113B (en) For the test strips and preparation method thereof of specificity tetanus antitoxin anaphylactogen IgE examination
CN103995133B (en) For the test strips and preparation method thereof of specificity egg white anaphylactogen IgE examination
CN110361543A (en) NTx Test paper, NTx detection kit and detection method

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
WW01 Invention patent application withdrawn after publication

Application publication date: 20210406

WW01 Invention patent application withdrawn after publication