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Enzyme-free signal amplification biomarker detection method based on surface-enhanced Raman spectroscopy

A surface-enhanced Raman and biomarker technology, which is applied in the field of chemical detection, can solve the problems of error in detection results, time-consuming enzymatic reaction, and high detection limit, and achieves the advantages of reducing detection cost, saving detection time, and reducing detection limit. Effect

Active Publication Date: 2021-08-03
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The disadvantages of this detection method are: the detection limit is relatively high, and the enzymatic reaction takes a long time
More importantly, the activity of the enzyme will change with the change of the external conditions. The fluctuation of temperature, pH and other conditions will have a greater impact on the enzyme-catalyzed reaction, which will cause greater errors in the detection results.

Method used

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  • Enzyme-free signal amplification biomarker detection method based on surface-enhanced Raman spectroscopy
  • Enzyme-free signal amplification biomarker detection method based on surface-enhanced Raman spectroscopy
  • Enzyme-free signal amplification biomarker detection method based on surface-enhanced Raman spectroscopy

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] A non-enzyme signal amplification biomarker detection method based on surface-enhanced Raman spectroscopy, comprising the following steps (see figure 1 ):

[0045] (1) Preparation of silver nanoparticles and modification of labeled antibodies

[0046] ① 40nm silver nanoparticles were synthesized by a one-pot method, and 18 mg of silver nitrate (AgNO 3 ) and 100mL ultrapure water, heated to slight boiling; dripping 2mL mass concentration of 1% trisodium citrate into the slightly boiling solution, continue heating and keep the solution boiling until the color of the solution remains unchanged, and then continue heating for 15min , obtaining silver nanoparticles (AgNPs) solution;

[0047] The absorption spectrum of the AgNPs solution was measured by UV-Vis absorption spectroscopy, and the measurement results can be found in figure 2 .

[0048] ② Centrifuge the silver nanoparticle solution at a high speed of 6000 r / min for 5 min, and then disperse it in a PBS solution ...

Embodiment 2

[0068] A method for detecting enzyme-free signal amplification biomarkers based on surface-enhanced Raman spectroscopy, comprising the following steps:

[0069] (1) Preparation of silver nanoparticles and modification of labeled antibodies

[0070] ① 20nm silver nanoparticles were synthesized by a one-pot method, and 9 mg of silver nitrate (AgNO 3 ) and 50mL ultrapure water, heated to slightly boiling; dropwise adding 1mL mass concentration of 1% trisodium citrate into the slightly boiling solution, continue heating and keep the solution boiling until the color of the solution remains unchanged, and then continue heating for 10min , obtaining silver nanoparticles (AgNPs) solution;

[0071] The absorption spectrum of the AgNPs solution was measured by UV-Vis absorption spectroscopy.

[0072] ② The silver nanoparticle solution was centrifuged at a high speed of 5500 r / min for 10 min, and then dispersed in a PBS solution with a concentration of 5 mM and pH=7.4.

[0073] ③ Add ...

Embodiment 3

[0092] A method for detecting enzyme-free signal amplification biomarkers based on surface-enhanced Raman spectroscopy, comprising the following steps:

[0093] (1) Preparation of silver nanoparticles and modification of labeled antibodies

[0094] ① 100nm silver nanoparticles were synthesized by a one-pot method, and 36 mg of silver nitrate (AgNO 3 ) and 200mL ultrapure water, heated to slightly boiling; dripping 3mL mass concentration of 1% trisodium citrate into the slightly boiling solution, continue heating and keep the solution boiling until the color of the solution remains unchanged, and then continue heating for 20min , obtaining silver nanoparticles (AgNPs) solution;

[0095] The absorption spectrum of the AgNPs solution was measured by UV-Vis absorption spectroscopy.

[0096] ② The silver nanoparticle solution was centrifuged at a high speed of 6500 r / min for 3 minutes, and then dispersed in a PBS solution with a concentration of 20 mM and pH=7.4.

[0097] ③ Add ...

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Abstract

The method for detecting enzyme-free signal amplification biomarkers based on surface-enhanced Raman spectroscopy of the present invention comprises the following steps: (1) preparing silver nanoparticles and modifying labeled antibodies; (2) constructing a sandwich immune sandwich structure; (3) preparing nano-gold and dual-functional surface enhancement Raman spectrum nanoprobe; (4) surface-enhanced Raman spectrum detection of biomarker standards and actual samples; the present invention uses metal Ag + Instead of enzymes as amplification means, the detection of enzyme-free signal amplification biomarkers based on surface-enhanced Raman spectroscopy was realized, and gold nanoparticles were co-modified with terminal alkynes and SERS probe molecules, and a nano-gold that can interact with Ag was obtained. + The dual-functional nanoprobe that responds and can obtain SERS signal improves the detection sensitivity and reduces the detection limit; replaces the traditional enzyme-catalyzed process with chemical reaction, which can save detection time, improve the stability of the detection system, and reduce the detection limit. The cost has positive significance for the introduction of chemical reactions into the biomarker detection system.

Description

technical field [0001] The invention relates to the technical field of chemical detection, and relates to the use of Ag + Aggregation of the double-functionalized gold nanoparticles results in an enhanced Raman signal for the probe molecule, and this principle is applied to the detection of biomarkers. Specifically, organic molecules containing terminal alkyne groups and surface-enhanced Raman probe molecules are used to double-functionalize gold nanoparticles. The organic molecules containing terminal alkyne groups and Ag + The reaction will cause the aggregation of bifunctional gold nanoparticles, so that the Raman signal obtained by the probe molecule is significantly enhanced, and this principle is applied to the detection of biomarkers. Background technique [0002] At present, the commonly used detection method of biomarkers is enzyme-linked immunosorbent assay (ELISA). The detection method is based on the specific combination of antigen and antibody, and the degree ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 马巍许多郭丹刘捷韩焕兴龙亿涛
Owner EAST CHINA UNIV OF SCI & TECH