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Immunobiosensor with prussian blue converted from magnetic beads for avian influenza virus and preparation method thereof

An avian influenza virus and biosensor technology, which can be applied to instruments, scientific instruments, electrochemical variables of materials, etc., can solve problems such as time-consuming and cumbersome, and achieve the effects of broad application prospects, sensitive detection, and simple modification operations.

Active Publication Date: 2016-06-15
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Other nanomaterials, such as noble metal nanoparticles, carbon nanotubes / fibers, and semiconductor quantum dots, are widely used for signal output due to their excellent surface modification functions and unique optical, electrical, and thermal properties. A large number of separation steps are involved in the process, and methods such as centrifugation, ultrafiltration and dialysis are frequently used, which is tedious and time-consuming

Method used

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  • Immunobiosensor with prussian blue converted from magnetic beads for avian influenza virus and preparation method thereof
  • Immunobiosensor with prussian blue converted from magnetic beads for avian influenza virus and preparation method thereof
  • Immunobiosensor with prussian blue converted from magnetic beads for avian influenza virus and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Put 15mL of magnetic bead aqueous solution in a 50mL centrifuge tube, and disperse for 5min under the action of ultrasound with a working frequency of 40kHz and a power of 160W to obtain a uniformly dispersed magnetic bead dispersion.

[0056] Then 15 mL of 0.4 M citric acid aqueous solution was added, ultrasonicated for 30 min, and rotated for 20 h at room temperature (25° C.). Perform magnetic separation, remove supernatant, and wash magnetic beads three times with phosphate buffer, then disperse in 15 mL MES buffer (final concentration of magnetic beads is about 1 mg / mL), and store at 4°C.

[0057] Add EDC (2mg, 10mM) and NHS (0.35mg, 15mM) to 1mL of the above functional magnetic bead dispersion, stir and react at room temperature for 2 hours, wash with phosphate buffer (pH6.0) magnetically for three times, and then disperse in 1mL phosphate buffer.

[0058] Take 100 μL of activated magnetic beads and 400 μL of concanavalin A (ConA) to react at room temperature for ...

Embodiment 2

[0067] Put 15mL of magnetic bead aqueous solution in a 50mL centrifuge tube, and disperse for 5min under the action of ultrasound with a working frequency of 40kHz and a power of 160W to obtain a uniformly dispersed magnetic bead dispersion.

[0068] Then add different volumes of 0.4M citric acid aqueous solution, so that the ratio of citric acid to magnetic bead dispersion is 0.1mmol, 0.2mmol and 0.3mmol per mg of magnetic nano material. Sonicate for 30 minutes, and rotate for 20 hours at room temperature (25° C.). Perform magnetic separation, remove supernatant, and wash magnetic beads three times with phosphate buffer, then disperse in 15 mL MES buffer (final concentration of magnetic beads is about 1 mg / mL), and store at 4°C.

[0069] Add EDC (2mg, 10mM) and NHS (0.35mg, 15mM) to 1mL of the above functional magnetic bead dispersion, stir and react at room temperature for 2 hours, wash with phosphate buffer (pH6.0) magnetically for three times, and then disperse in 1mL pho...

Embodiment 3

[0074] Put 15mL of magnetic bead aqueous solution in a 50mL centrifuge tube, and disperse for 5min under the action of ultrasound with a working frequency of 40kHz and a power of 160W to obtain a uniformly dispersed magnetic bead dispersion.

[0075] Then 15 mL of 0.4 M citric acid aqueous solution was added, ultrasonicated for 30 min, and rotated for 20 h at room temperature (25° C.). Perform magnetic separation, remove supernatant, and wash magnetic beads three times with phosphate buffer, then disperse in 15 mL MES buffer (final concentration of magnetic beads is about 1 mg / mL), and store at 4°C.

[0076] Add EDC (2mg, 10mM) and NHS (0.35mg, 15mM) to 1mL of the above functional magnetic bead dispersion, stir and react at room temperature for 2 hours, wash with phosphate buffer (pH6.0) magnetically for three times, and then disperse in 1mL phosphate buffer.

[0077] Take 100 μL of activated magnetic beads and 400 μL of concanavalin A (ConA) to react at room temperature for ...

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PUM

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Abstract

The invention discloses an immunobiosensor with prussian blue converted from magnetic beads for avian influenza virus and a preparation method thereof. An electrode modified with antibodies is used for capturing avian influenza virus H5N1, and magnetic beads are marked; under the control of a magnetic field, an electrochemical method is used for converting magnetic bead markers to generate prussian blue; an electrochemical reduction peak based on the generated prussian blue is used for quantitative determination of avian influenza virus, and the biosensor is formed. A new property of magnetic nanometer particles is provided innovatively, prussian blue with electrochemical activity is generated by introduced electrochemical conversion based on fully utilization of separating and enriching functions of magnetic beads, so that the biosensor for sensitive detection of avian influenza virus H5N1 is obtained; the biosensor has the advantages of simple methods, low cost, and good application prospect, and can be used for sensitively detecting avian influenza virus H5N1.

Description

technical field [0001] The invention relates to the fields of nanometer material science and electrochemical biosensing, in particular to an avian influenza virus immune biosensor and method for converting magnetic beads into Prussian blue. Background technique [0002] As a kind of nanomaterials, magnetic nanomaterials not only have the advantages of large specific surface area, good biocompatibility, rich surface functional groups, and high surface free energy, they are widely used to efficiently immobilize a large number of biomolecules, and based on their unique magnetic properties in magnetic Separation and enrichment and magnetic hyperthermia and other fields have received much attention. However, the current applications of magnetic beads are mainly based on magnetism, and the application of their own chemical / electrochemical properties is rarely developed. [0003] In biosensing research, signal output and amplification are one of the most critical factors determini...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N33/569
CPCG01N27/26G01N33/56983
Inventor 傅迎春李玲艳李延斌
Owner ZHEJIANG UNIV
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