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Preparation method of electrochemical sensor with surface curing polypeptide probe

A surface curing and electrochemical technology, applied in scientific instruments, instruments, material analysis by electromagnetic means, etc., can solve the requirements of extremely high surface flatness of metal electrodes, change the third- and fourth-order structures of polypeptides, and be unfavorable for large-scale safety. Production and other issues, to achieve the effect of retaining the effective activity and sensor sensitivity, simplifying the production process and cost, and increasing the maximum withstand current density and sensitivity

Active Publication Date: 2017-01-04
MICRO TECH MEDICAL HANGZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are deficiencies in the existing curing processes of various probe molecules, which are mainly reflected in: the peptide cross-linking process usually uses glutaraldehyde as a cross-linking medium to promote covalent cross-linking between polypeptide molecules and adjacent molecules. It will change the third-order and fourth-order structures of polypeptides, significantly reducing the activity and sensitivity of probe molecules, especially enzyme molecules; and glutaraldehyde is an extremely flammable and volatile toxic substance, which is not conducive to safe production in large quantities
Its disadvantage is that it has extremely high requirements on the flatness of the metal electrode surface, and can only achieve single-layer curing, so the molecular density of the cured probe is low, and the sensitivity performance of the sensor is poor.
The disadvantage is that most photosensitive adhesives have certain biological toxicity, which is not suitable for use in in vivo sensors, and the precision ultraviolet exposure equipment is expensive, and the development process is more complicated, making its manufacturing cost higher
Among them, the glue layer needs to meet the two requirements of biocompatibility and the ability to bond two layers of substances with completely different physical and chemical properties, so the options are very limited, and often need to be developed separately according to the specific properties of the probe molecular layer and the permeable membrane layer. lead to high manufacturing costs
In the chemical reaction process, due to the high sensitivity of the polypeptide probe molecules to reaction conditions such as temperature, pH of the solution, and organic or corrosive solvents, in order to ensure that the probe molecules will not be destroyed, the practical chemical reaction options are also very limited.

Method used

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  • Preparation method of electrochemical sensor with surface curing polypeptide probe
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preparation example Construction

[0033] A method for preparing an electrochemical sensor with a surface-cured polypeptide probe, comprising the steps of:

[0034] Step 1. The electrodes of the electrochemical sensor are prepared on the substrate by conventional electrode processing technology; the conventional electrode processing technology includes metal coating etching or conductive paste screen printing process, and the substrate includes inorganic substrates such as glass, silicon wafers, etc. or organic substrates. The substrate is polyimide, polydimethylsiloxane, etc., wherein the working electrode is carbon, graphene, gold or platinum.

[0035] Step 2, utilize oxidized dopamine to carry out cross-linking nanometer metal particle and polypeptide probe molecule, and be attached on the working electrode, promptly form polydopamine probe layer on the working electrode; Polypeptide probe molecule comprises enzyme protein (such as glucose oxidase, Glucose dehydrogenase, horseradish peroxidase and other oxid...

Embodiment 1

[0043] A method for preparing an electrochemical sensor with a surface-cured polypeptide probe, comprising the steps of:

[0044] Step 1. Prepare the electrode of the electrochemical sensor on the glass substrate by metal coating etching process, wherein the working electrode is platinum.

[0045] Step 2, adding polypeptide probe molecule (glucose oxidase), dopamine molecule and chloroplatinic acid into water, the mass concentration of polypeptide probe molecule is 5mg / mL, the mass concentration of dopamine molecule is 5g / L, the mass concentration of chloroplatinic acid The mass concentration is 5mg / L; the pH is 8, the dissolved oxygen saturation is lower than 1%, and the reaction is carried out at 30°C to generate surface-cured oxidized polydopamine and nano-metal particles of polypeptide probe molecules; The substrate is put into the mixed solution, and the nanometer metal particles are deposited on the surface of the working electrode by electrochemical oxidation reaction. ...

Embodiment 2

[0049] A method for preparing an electrochemical sensor with a surface-cured polypeptide probe, comprising the steps of:

[0050] Step 1, using a conductive paste screen printing process to prepare electrodes for electrochemical sensors on polydimethylsiloxane substrates, wherein the working electrodes are gold.

[0051] Step 2, adding gold nanoparticles, polypeptide probe molecules (hepatitis B virus antibody) and dopamine molecules into water, the size of gold nanoparticles is about 50 nanometers, the mass concentration of gold nanoparticles is 25000ppm, and the mass concentration of polypeptide probe molecules is 4mg / mL, and the mass concentration of dopamine molecules is 6g / L; and the substrate in step 1 is immersed, and the pH is 7, the dissolved oxygen saturation is lower than 1%, 35°C, and the voltage applied on the working electrode is relative to the solution The silver / silver chloride reference electrode in the method performs electrochemical oxidation reaction under...

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Abstract

The invention discloses a preparation method of an electrochemical sensor with a surface curing polypeptide probe. The method comprises the following steps that electrodes of the electrochemical sensor are prepared on a base material; oxidized dopamine is utilized for cross-linking metal nanometer particles and polypeptide probe molecules, and the cross-linked metal nanometer particles and polypeptide probe molecules are attached to a working electrode, that is to say, a polydopamine probe layer is formed on the working electrode; a permeable membrane solution is plated to the polydopamine probe layer and dried, and a permeable membrane solution is formed. According to the preparation method of the electrochemical sensor with the surface curing polypeptide probe, the multiple purposes such as electrode modification, polypeptide probe molecule curing and bonding layer adding are achieved in the same process simultaneously through a dopamine oxidation polymerization reaction, the manufacturing process is greatly simplified, and the manufacturing cost is reduced. Dopamine has good biocompatibility, achieves an excellent adhesive effect and can be closely boned to almost all types of surfaces. Therefore, for the manufacturing process, the step of adding biocompatibility adhesives or conducting bonding treatment after permeable membrane coating can be omitted, the production process is simplified, and the cost is reduced.

Description

technical field [0001] The invention relates to the technical field of electrochemical sensors, in particular to a method for preparing an electrochemical sensor with a surface-cured polypeptide probe. Background technique [0002] An electrochemical sensor is a sensor that detects the content or concentration of a target analyte in a sample with an electrochemical signal, which is generated by a change in Faradaic current or a change in properties such as AC impedance. Generally, an electrochemical sensor is composed of at least one working electrode, a counter electrode and a reference electrode, and the working electrode has probe molecules that can bind or chemically react with the target analyte in the detection sample. [0003] Currently available probe molecules include ribonucleic acid chains, enzymes, polypeptide antibodies or other proteins that can bind to analytes, etc. Among them, proteins such as enzymes and antibodies can be collectively referred to as polypep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3278
Inventor 于非
Owner MICRO TECH MEDICAL HANGZHOU CO LTD
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