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A method for protein detection based on nano-gold-dna composites

A composite material and nano-gold technology, applied in the field of protein detection, can solve the problems of complex operation, heavy workload, and difficulty in realizing high-throughput rapid detection of proteins, and achieve the effect of cumbersome preparation process

Inactive Publication Date: 2018-03-20
CAPITAL NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is not only complicated in operation and heavy in workload, antigens and antibodies must come from living organisms, resulting in extremely high economic costs, but also their types are limited
Both of these methods are difficult to achieve high-throughput rapid detection of proteins

Method used

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  • A method for protein detection based on nano-gold-dna composites
  • A method for protein detection based on nano-gold-dna composites
  • A method for protein detection based on nano-gold-dna composites

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Method for detecting protein based on nano-gold-DNA composite material

[0036] The method for detecting protein based on nano-gold-DNA composite material provided in this embodiment includes the following steps:

[0037] 1. Preparation of gold nanoparticles: prepared by reducing chloroauric acid with sodium citrate, the concentration of chloroauric acid solution is 50mM, the molar ratio of chloroauric acid and sodium citrate is 3.5:1, and the average gold nanoparticles in the resulting solution The particle size is about 13nm;

[0038] 2. Preparation of gold nano-DNA complex: add 10 μL of a DNA solution with a concentration of 1 μM at one end modified with sulfhydryl group to 50 μL of the above solution containing gold nanoparticles. The DNA is poly(A) 15 , Get nano gold-poly(A) 15 Complex;

[0039] 3. Add 50μL of each of the 11 protein solutions with a concentration of 300nM (100nM or 500nM) to 60μL of the above solution containing the gold nano-DNA complex, and inc...

Embodiment 2

[0049] Example 2 Detection of protein mixtures of different components based on nano-gold-DNA composite materials

[0050] It includes the following steps:

[0051] 1. Preparation of gold nanoparticles: prepared by reducing chloroauric acid with sodium citrate, the concentration of chloroauric acid solution is 50mM, the molar ratio of chloroauric acid and sodium citrate is 3.5:1, and the average gold nanoparticles in the resulting solution The particle size is about 13;

[0052] 2. Preparation of gold nano-DNA complex: add 10 μL of a DNA solution with a concentration of 1 μM at one end modified with sulfhydryl group to 50 μL of the above solution containing gold nanoparticles. The DNA is poly(A) 15 , Get nano gold-poly(A) 15 Complex;

[0053] 3. Add 50 μL each of the two protein mixed solutions (BSA and HSA mixed solution or BSA and Con mixed solution) with a concentration of 300 nM to 60 μL of the above solution containing the gold nano-DNA complex, and incubate each at 37°C for 30 m...

Embodiment 3

[0063] Example 3 Identifying proteins in serum samples based on nano-gold-DNA composite materials

[0064] It includes the following steps:

[0065] 1. Preparation of gold nanoparticles: prepared by reducing chloroauric acid with sodium citrate, the concentration of chloroauric acid solution is 50mM, the molar ratio of chloroauric acid and sodium citrate is 3.5:1, and the average gold nanoparticles in the resulting solution The particle size is about 13;

[0066] 2. Preparation of gold nano-DNA complex: add 10 μL of a DNA solution with a concentration of 1 μM at one end modified with sulfhydryl group to 50 μL of the above solution containing gold nanoparticles. The DNA is poly(A) 15 , Get nano gold-poly(A) 15 Complex;

[0067] 3. Add 50μL of each of the 11 protein solutions with a concentration of 3μM to 60μL of the above solution containing gold nano-DNA complex, and incubate at 37°C for 30min; do at least 5 replicates for each protein; 2 O is a blank control, that is, add ddH to 60 ...

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Abstract

The invention provides a method for detecting protein based on a nanogold-DNA composite material, comprising the steps: preparing the nanogold-DNA composite, producing a nanogold-DNA-protein ternary composite, catalyzing nitrophenol and sodium borohydride with nanogold for reaction (a color development system), visually inspecting the period of time during which the color of reaction liquid changes from yellow to colorless (CCT value), drawing an LDA chart according to the CCT / CCT0 values of different protein, adding protein with given concentration to be detected into the color development system, calculating the CCT / CCT0 values of the protein to be detected, and identifying the protein to be detected with combination of the LDA chart. The method is established on the basis of a linear distinguishing and analysis technology and can successfully identify different protein only through visually inspecting and recording with a timer the period of time during which the color of the liquid changes, no expensive instrument is needed, the detection limit achieves 30nM, and a new technological means is provided for high flux quick detection of protein.

Description

Technical field [0001] The invention relates to a method for detecting proteins based on nano-gold-DNA composite materials. Background technique [0002] Protein detection provides valuable information for the early diagnosis of many diseases, such as hypoalbuminemia, Alzheimer’s disease, cancer, prostate inflammation, AIDS, etc. (U.Andreasson; E. Portelius; ME Andersson, et al. .,Biomarker Med.2007,1,59-78;VUBai;A.Kaseb;S.Tejwani,etal.,Proc.Natl.Acad.Sci.2007,104,2343-2348;J.Hardy;DJSelkoe, Science 2002, 297, 353-356). At present, protein detection methods mainly include two categories: one is high performance liquid chromatography-mass spectrometry and two-dimensional gel electrophoresis. This method has the disadvantages of requiring professional operation, expensive equipment, and time-consuming. The other is an immunization method based on antigen-antibody specific recognition of biomarkers. This method is not only complicated to operate, heavy workload, antigens and ant...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/532
CPCG01N33/532G01N33/6803
Inventor 陈郑博魏祥聪谭路路
Owner CAPITAL NORMAL UNIVERSITY
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