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Nanogold biochip for detecting Alzheimer's disease marker as well as preparation method and application of nanogold biochip

An Alzheimer's disease, biochip technology, applied in measurement devices, material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of not reaching the picogram level, inaccurate results, low sensitivity, etc., and improve the emission intensity. , the effect of improving the detection signal and improving the sensitivity

Pending Publication Date: 2022-03-01
SHENZHEN UNIV +1
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Among these research methods, the results of the colorimetric method are intuitively visible to the naked eye, but the sensitivity is low; although the sensitivity of the fluorescence method is slightly higher than that of the colorimetric method, it still cannot reach the picogram level, and it is susceptible to blood contamination when detecting real blood samples. The interference of autofluorescence makes the results inaccurate; the sensitivity of electrochemiluminescence is the highest among several methods, but electrochemiluminescence requires professional detection instruments and is expensive

Method used

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  • Nanogold biochip for detecting Alzheimer's disease marker as well as preparation method and application of nanogold biochip
  • Nanogold biochip for detecting Alzheimer's disease marker as well as preparation method and application of nanogold biochip
  • Nanogold biochip for detecting Alzheimer's disease marker as well as preparation method and application of nanogold biochip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Preparation of Aβ40 nano-gold biochip and protein detection

[0081] 1. Preparation of nano-gold biochip

[0082] (1) Depositing gold nanoparticles by wet method on a glass substrate to obtain a nano-gold chip;

[0083] (2) Print the capture antibody of Aβ40 with a concentration of 1.0 μg / mL on the nano-gold chip by using a micro-spotter;

[0084] (3) Further print the chicken IgY antibody on the chip as an internal reference control to obtain a nano-gold biochip;

[0085] 2. Coupling of capture antibody and fluorescent molecule IRDye800

[0086] (4) Mix the capture antibody of Aβ40 and the fluorescent molecule IRDye800 according to the molar ratio of 1:10, react for half an hour, then mix evenly, and continue to react for half an hour;

[0087] (5) After the reaction is completed, add an appropriate amount of PBS to make the volume of the reaction system reach 500 μl;

[0088] (6) Add 10 mL of PBS to the NAP-5 column, wash the column, and when the solution in the c...

Embodiment 2

[0102] Preparation of Aβ42 nano-gold biochip and protein detection

[0103] The difference between the preparation of the Aβ42 nano-gold biochip and the preparation method of the Aβ40 nano-gold biochip in Example 1 is that the capture antibody of Aβ42 is printed on the nano-gold chip to obtain the Aβ42 nano-gold biochip;

[0104] The difference between the detection of Aβ42 protein and the detection of Aβ40 protein in Example 1 is that the standard curve of Aβ42 is obtained by using the Aβ42 protein standard, and the protein to be tested is Aβ42 protein.

[0105] Fluorescent images of Aβ42 and standard curves such as Figure 5 Shown, where, y=0.55x+0.34, R 2 = 0.994.

[0106] In summary, the present invention proposes a method for preparing a nano-gold biochip for detecting Alzheimer's disease markers. Gold nanoparticles are deposited on the chip substrate, so that the surface of the chip is distributed with many discontinuous, Gold nanoparticles with uniform spacing, and t...

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Abstract

The invention provides a nanogold biochip for detecting an Alzheimer's disease marker and a preparation method and application of the nanogold biochip. Gold nanoparticles are deposited on a chip substrate, so that a plurality of discontinuous gold nanoparticles with uniform intervals are distributed on the surface of the chip; and fixing a capture antibody for detecting the Alzheimer's disease marker on the chip through printing to form a capture layer, thereby obtaining the nanogold biochip. A local electric field is generated due to gaps of the gold nanoparticles, so that the emission intensity of the coupled fluorescent molecules is improved, surface plasmas and the fluorescent molecules resonate, and fluorescence detection signal amplification is realized. The gold nanoparticles can also increase the specific surface area and increase binding sites for capturing the antibody. The near-infrared fluorescent molecules are adopted, so that background fluorescent signals of the substrate can be effectively avoided, and the signal-to-noise ratio is increased. Finally, background noise is reduced by detecting near-infrared fluorescence, a fluorescence output signal is enhanced by the gold nanoparticles, and the sensitivity of Alzheimer's disease biomarker detection is improved.

Description

technical field [0001] The invention relates to the field of biochemical detection, in particular to a nano-gold biochip for detecting Alzheimer's disease markers, a preparation method and application thereof. Background technique [0002] Alzheimer's disease (AD) is a complex neurodegenerative disease with a very long duration. The detection of blood markers is of great significance for the early diagnosis and universal screening of AD. However, the content of AD blood markers is low, which has exceeded the detection range of traditional detection methods (ELISA). Therefore, it is necessary to develop highly sensitive and specific detection methods to achieve ultra-sensitive detection of AD blood markers. [0003] Researchers have established colorimetric methods, fluorescence methods, electrochemical methods, electrochemiluminescence methods and other methods for detection. Among these research methods, the results of the colorimetric method are intuitively visible to the...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N21/01
CPCG01N21/6486G01N21/01
Inventor 徐庆刘一镳刘琼王国新廖滔陈敏文
Owner SHENZHEN UNIV
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