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Preparation method of a biosensor for sensitive detection of protein

A biosensor and sensitive detection technology, applied in the fields of electrochemistry and nanomaterials, can solve the problems of large specific surface area, small particle size of nano-silver particles, high surface reactivity, etc., and achieve simple operation, strong repeatability, and enhanced sensitivity Effect

Active Publication Date: 2019-09-03
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the small particle size, large specific surface area, and high surface reactivity of silver nanoparticles, they have good catalytic activity, but the use of silver nanomaterials as electrode modifiers for the determination of proteins has not been reported at home and abroad.

Method used

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  • Preparation method of a biosensor for sensitive detection of protein
  • Preparation method of a biosensor for sensitive detection of protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1. Preparation of surface functionalized polystyrene microspheres:

[0025] (1) Add 17.6mL of absolute ethanol, 20.8mL of deionized water and 2.1g of polyvinylpyrrolidone (PVP) into a three-necked flask equipped with a condenser, and place it in a constant temperature water bath at 70°C, mechanically stir and keep The rotating speed is 315r / min.

[0026] (2) Dissolve 2.1 g of initiator AIBN in 6 mL of styrene to obtain monomeric styrene in which the initiator AIBN is dissolved.

[0027] (3) Add styrene monomer dissolved in initiator AIBN to the system obtained in (1), and add 6 mL of acrolein (C 3 h 4 O), after a constant temperature reaction for 7 hours, add hydrochloric acid and raise the temperature of the constant temperature water bath to 80°C for 12 hours. After stopping the reaction, wash with ethanol / water and ultrasonic several times to obtain surface functionalized polystyrene microspheres (PS- CHO) emulsion.

[0028] 2. Preparation of surface-modified sil...

Embodiment 2、3、4、5、6

[0038] The same steps as in Example 1 will not be repeated here, only the concentration of the BSA protein solution prepared in step 3 is changed: the BSA protein is dispersed in 10 mM phosphate buffered saline solution (PBS, pH 7.2), and 10 -14 mol / L, 10 -13 mol / L, 10 -12 mol / L, 10 -11 mol / L and 10 -10 mol / L BSA solution.

[0039] Other steps are the same as in Example 1.

[0040] 2. Analysis of test results:

[0041] Electrochemical experiments were carried out on a CH1660D electrochemical workstation with a three-electrode system: the working electrode was a self-made modified conductive glass, the reference electrode was a saturated calomel electrode, and the counter electrode was a platinum wire electrode. The experimental temperature was 25°C.

[0042] Cyclic voltammetry test method: first configure a 0.1mol / L PBS buffer solution with a pH of 7.2, add 10mL of the prepared PBS buffer solution into the electrolytic cell, and then insert a saturated calomel electrode ...

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Abstract

The invention relates to a preparation method of a biosensor for sensitive detection of protein, which belongs to the fields of nanomaterials and electrochemistry. The invention adopts a self-assembly method to plate a layer of silver film on the conductive surface of ITO conductive glass, and further manufactures a biosensor electrode, which is studied and used for the detection of BSA. Using silver nanoparticles as markers, the self-conductivity of silver nanoparticles is used for detection by direct detection electrochemical method. The preparation method of the biosensor for the sensitive detection of proteins is low in production cost, strong in repeatability, does not need signal amplification during detection at the same time, has simple detection steps, and does not require complicated operations.

Description

technical field [0001] The invention belongs to the fields of nanometer material science and electrochemistry. Background technique [0002] In recent years, in the field of bioelectrochemistry. Direct electrochemical studies of electron transfer between proteins and electrodes have received increasing attention. Due to the extended three-dimensional structure of the protein, its electroactive center is embedded in the polypeptide chain, and its adsorption and denaturation on the surface of the conventional electrode will cause the passivation of the electrode surface, so the electron transfer speed of the protein on the general electrode is very slow. An effective current response cannot be obtained. [0003] Nanomaterials are one of the current hot research fields. Silver nanoparticles have excellent electrical conductivity and electrocatalytic properties. The preparation of silver nanoparticles modified electrodes can be used as organic memory, biosensors, capacitors, c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
Inventor 严长浩严欣刘康宇
Owner YANGZHOU UNIV
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