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Surface-enhanced Raman immunoassay planar sensor as well as preparation method and application thereof

A surface-enhanced Raman and immunoassay technology, applied in the fields of biosensing and medical diagnosis, can solve the problems of sample loss and cumbersome steps, and achieve the effect of sensitive detection and convenient and quick operation.

Pending Publication Date: 2020-12-01
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The above methods all use dispersed nanoparticles as the SERS substrate and SERS label. During the detection process, after the analyte is combined with the substrate and SERS label again, the nanoparticles need to be separated and separated by repeated centrifugation or introducing a magnetic field. Washing, cumbersome steps, easy to cause sample loss

Method used

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  • Surface-enhanced Raman immunoassay planar sensor as well as preparation method and application thereof
  • Surface-enhanced Raman immunoassay planar sensor as well as preparation method and application thereof
  • Surface-enhanced Raman immunoassay planar sensor as well as preparation method and application thereof

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Embodiment 1

[0055] 1) Preparation of SERS planar substrate:

[0056] The nano-gold particle solution with an average particle diameter of 10nm is centrifuged, and the precipitate is collected and diluted to 1.5mmol / L with ultrapure water, and cyclohexane; -Water two-phase interface, and then ethanol is quickly added to the two-phase solution at a volume ratio of ethanol:nano-gold solution=1:1, so that the nano-gold particles form a neatly arranged array at the two-phase interface. After the upper layer of cyclohexane volatilizes, an ordered nano-gold array is formed at the water-air two-phase interface. Insert a clean and dry glass slide below the liquid surface of the gold nano array at an angle, and quickly stir up the glass slide to generate a SERS substrate loaded with the gold nano array. After natural drying, soak the SERS substrate in 1mg / mL dopamine hydrochloride solution and incubate for 12h, wash with ultrapure water, then immerse in 20mmol / L, pH9.0 Tris-HCl buffer, add 100μg / m...

Embodiment 2

[0063] 1) Preparation of SERS planar substrate:

[0064] The nano-gold particle solution with an average particle diameter of 20nm is centrifuged, the precipitate is collected and diluted to 1.3mmol / L with ultrapure water, and ethyl acetate is added thereto at a volume ratio of ethyl acetate:nano-gold solution=1:1 to form an organic -Water two-phase interface, and then ethanol is quickly added to the two-phase solution at a volume ratio of ethanol:nano-gold solution=1:4, so that the nano-gold particles form a neatly arranged array at the two-phase interface. After the upper layer of ethyl acetate volatilized, an ordered nano-gold array was formed at the water-air two-phase interface. Insert a clean and dry glass slide below the liquid surface of the gold nano array at an angle, and quickly stir up the glass slide to generate a SERS substrate loaded with the gold nano array. After natural drying, soak the SERS substrate in 20mg / mL dopamine hydrochloride solution and incubate f...

Embodiment 3

[0071] 1) Preparation of SERS planar substrate:

[0072]The nano-gold particle solution with an average particle diameter of 50nm is centrifuged, the precipitate is collected and diluted to 1mmol / L with ultrapure water, and dichloromethane is added thereto at a volume ratio of dichloromethane: nano-gold solution=1:4 to form an organic- Water two-phase interface, and then ethanol is quickly added to the two-phase solution at a volume ratio of ethanol: gold nanometer solution = 1:3, so that the gold nanoparticles form a neatly arranged array at the two-phase interface. After the upper layer of dichloromethane volatilizes, an ordered nano-gold array is formed at the water-air two-phase interface. Insert a clean and dry glass slide below the liquid surface of the gold nano array at an angle, and quickly stir up the glass slide to generate a SERS substrate loaded with the gold nano array. After natural drying, soak the SERS substrate in 10mg / mL dopamine hydrochloride solution and ...

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Abstract

The invention relates to a surface-enhanced Raman immunoassay planar sensor as well as a preparation method and application thereof. The sensor comprises an SERS planar substrate and an SERS label, the preparation method of the sensor is applied to molecular detection of disease markers. The SERS planar substrate is prepared by modifying dopamine, a to-be-detected object antibody and bovine serumalbumin on the surface of a two-dimensional nanogold array. The SERS label is prepared by modifying report molecules on the surfaces of gold nanoparticles, packaging bovine serum albumin, and modifying dopamine and the to-be-detected object antibody. The sensor has the advantages that (1) in sample detection, the planar sensor is cleaned through leaching, so that convenience and quickness are realized; (2) the ordered nanogold array generates uniform hot spots, and the signal reproducibility is good; (3) the Raman detection sensitivity can be enhanced through the approaching of the SERS substrate and the label; and (4) the specificity of the immune sandwich method is strong, qualitative and quantitative detection of target molecules can be realized by combining the immune sandwich method with SERS, the response is sensitive, and the detection time is short.

Description

technical field [0001] The invention relates to a method for preparing and using a Surface-Enhanced Raman Scattering Immunoassay (Surface-Enhanced Raman Scattering Immunoassay) sensor, belonging to the fields of biosensing and medical diagnosis. Background technique [0002] Raman spectroscopy (Raman Spectroscopy) technology is a spectral analysis detection technology developed based on the inelastic scattering effect of light. Surface Enhanced Raman Scattering (SERS) is a phenomenon in which the Raman signal of the molecule is greatly enhanced when the molecule of the substance to be measured is adsorbed on the surface of metal sol particles (such as gold, silver, copper) or rough metal surface , has been effectively developed in food safety and environmental pollution detection. However, SERS technology still faces great challenges in medical diagnosis, because usually the concentration of biomarker molecules in disease diagnosis is low, and the components in serum or uri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65G01N33/532G01N33/543G01N33/552G01N33/577G01N33/58
CPCG01N21/658G01N33/587G01N33/577G01N33/552G01N33/54366G01N33/532
Inventor 王梦凡屈琪王静曾川齐崴
Owner TIANJIN UNIV
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