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Biodetector based on interference effect of ordered porous nanostructured thin film and method for detecting biomolecules

A thin film interference and nanostructure technology, which is applied in nanoparticle analysis, biological particle analysis, suspension and porous material analysis, etc. It can solve the problems of limited biological molecular weight, limited amplitude of interference spectrum, and reduced detection sensitivity, and achieve the preparation process. Cumbersome, good commercialization prospects, and the effect of improving sensitivity

Active Publication Date: 2020-07-31
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the disadvantages of this planar interference layer are obvious: firstly, the cost of this coating is expensive, which makes the detection cost high; secondly, the specific surface area of ​​the planar interference layer is limited, and the biological molecular weight that can be captured is limited, so the detection sensitivity is often up to less than the requirements; finally, the amplitude of the interference spectrum of this type of interference layer is also very limited, and the signal-to-noise ratio is low, which will also reduce the detection sensitivity to a certain extent

Method used

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  • Biodetector based on interference effect of ordered porous nanostructured thin film and method for detecting biomolecules
  • Biodetector based on interference effect of ordered porous nanostructured thin film and method for detecting biomolecules
  • Biodetector based on interference effect of ordered porous nanostructured thin film and method for detecting biomolecules

Examples

Experimental program
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Effect test

Embodiment 1

[0096] The concentration of hepatitis B surface antigen (HBsAg) in the solution is detected by the biodetector of the present invention, wherein the sensing unit is an ordered porous polystyrene film with three-dimensional ordered nanopores, and the pore size is 200nm. Specific steps are as follows:

[0097] 1) Modification of the sensing unit (antibody coating):

[0098] Take a polytetrafluoroethylene reaction cell 17 with an inner diameter of about 7 mm, seal the edge with a rubber ring 18, and fix the polytetrafluoroethylene reaction cell 17, rubber ring 18, sensor sheet 19 and slide glass 20 under pressure with a small stationery clip Together, as shown in Figure 4. The sensing sheet 3 is an ordered porous polystyrene film. Inject 200 μl of coating solution into the above-mentioned polytetrafluoroethylene reaction cell, and the coating solution is a solution obtained by dissolving Anti-HBs monoclonal antibody in 0.05M carbonate buffer solution (CB buffer solution) of pH ...

Embodiment 2

[0107] Use the biodetector of the present invention to observe the covalent coupling between glutaraldehyde (Glutaraldehyde, GA) and IgG (ie antibody) in human serum, wherein the sensing unit is an ordered porous with three-dimensional ordered nanopores Polystyrene film with a pore size of 200nm. Specific steps are as follows:

[0108]1) Fix the ordered porous polystyrene film sensor sheet with three-dimensional ordered nanopores in the detection cell, inject a certain amount of absolute ethanol into the detection cell to fill the detection cell; PBS buffer was injected into the detection cell at a rate of 200 μl / min, and the optical thickness of the ordered porous polystyrene film sensor sheet was recorded as a function of time until it stabilized;

[0109] 2) Prepare glutaraldehyde solution (glutaraldehyde solution is prepared by pH 5.0, 0.05M PBS buffer solution, and the concentration of glutaraldehyde in the glutaraldehyde solution is 200mM), and the glutaraldehyde soluti...

Embodiment 3

[0113] Utilize the biodetector of the present invention to observe the interaction between staphylococcal protein A (SPA) and IgG (antibody) in human serum, wherein the sensing unit is an ordered porous polystyrene film with three-dimensional ordered nanopores, The pore size is 200nm. Specific steps are as follows:

[0114] 1) Fix the ordered porous polystyrene film sensor sheet with three-dimensional ordered nanopores in the detection cell, inject a certain amount of absolute ethanol into the detection cell to fill the detection cell; PBS buffer was injected into the detection cell at a rate of 200 μl / min, and the optical thickness of the ordered porous polystyrene film sensor sheet was recorded as a function of time until it stabilized;

[0115] 2) The SPA solution (the SPA solution is prepared by pH 9.6, 0.05M CB buffer solution, the SPA concentration in the SPA solution is 5 μg / ml) is injected into the detection cell at a flow rate of 200 μl / min, and the ordered porous po...

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Abstract

The invention discloses a biological detector based on the interference effect of an ordered porous nanostructure film. The biological detector includes a detection pool, a sensing unit arranged in the detection pool, and an optical fiber spectrometer for detecting the reflection interference spectrum of the sensing unit. The sensing unit is a porous film with three-dimensional ordered nanopores, and the diameter of the nanopores is 20nm-500nm. The invention also discloses a method for detecting biomolecules by using the biodetector. When the substance to be measured is adsorbed on the pore surface of the ordered porous film as the sensing unit, it will change the spectral distribution of the reflected interference light, resulting in a spectral line shift, which can be used to quantitatively or qualitatively analyze the sample The concentration of substances in the medium, the attachment rate, the interaction, the change of the geometric size, etc.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a biological detection instrument based on the interference effect of ordered porous nanostructure thin films and a method for detecting biological molecules. Background technique [0002] A biosensor is an instrument that is sensitive to biological substances and converts its concentration into electrical signals for detection. It uses immobilized biosensitive materials as recognition elements (including enzymes, antibodies, antigens, microorganisms, cells, tissues, Analytical tools or systems composed of nucleic acid and other biologically active substances), appropriate physical and chemical transducers (such as oxygen electrodes, photosensitive tubes, field effect tubes, piezoelectric crystals, etc.) and signal amplification devices. Biosensors have the functions of receptors and converters. The application of biosensors involves many aspects, one of the main ap...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/55
CPCG01N21/55G01N15/0205G01N2015/0693G01N2015/0038G01B11/06G01N15/06G01N2015/1006G01N2015/0065G01N1/14
Inventor 钱卫平苏倩倩钱辰
Owner SOUTHEAST UNIV
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