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Method for quantitatively detecting protein through quantum dot resonant scattering

A quantitative detection and quantum dot technology, which is applied in the field of protein detection, can solve the problem of unquantified detection of proteins, etc., and achieve the effect of high sensitivity and good stability.

Inactive Publication Date: 2012-07-11
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In the reports of the existing literature, although there are organic dyes or metal ion indicators for the analysis of the resonance scattering spectroscopic analysis of the binding of biomacromolecular proteins and nucleic acids, and the analysis of the resonance scattering characteristics of quantum dots, so far no use of quantum dots has been made. Literature Reports on Resonance Scattering for Quantitative Detection of Proteins

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  • Method for quantitatively detecting protein through quantum dot resonant scattering
  • Method for quantitatively detecting protein through quantum dot resonant scattering
  • Method for quantitatively detecting protein through quantum dot resonant scattering

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Embodiment 1

[0037] 1) Preparation of ZnS / ZnO quantum dot heterostructure: First, 2mmol Zn(Ac) 2 and 1.4 mmol CH 3 CSNH 2 Add 20ml of 1,4-butanediol, and ultrasonically disperse until a transparent precursor solution is formed; then transfer the obtained solution to a 30mL hydrothermal reaction kettle (lined with polytetrafluoroethylene), and react at 200°C for 18h; After the thermal reaction kettle was naturally cooled to room temperature, 10-20ml of distilled water was added to the obtained product to mix, centrifuged at 8000rpm, the obtained precipitate was washed with ethanol three times to remove impurities, and dried at 60°C for 5 hours to obtain the ZnS / ZnO quantum dot heterostructure. The diameter is 3~4nm.

[0038] 2) Silicon-coating step: Dissolve 0.1mmol ZnS / ZnO quantum dots in 40mL ethanol, add 0.5mL 28wt% ammonia water and mix well, then slowly add 0.1mL TEOS under continuous stirring, and continue stirring for 12 hours. Centrifuge to get precipitate, wash twice with distil...

Embodiment 2

[0041] Embodiment 2 takes the detection of bovine serum albumin (BSA) as an example

[0042] (1) Preparation of quantum dot solution and protein solution

[0043] The original concentration of quantum dots 1 to 4 is 5×10 -5 mol / L, diluted 60 times with double distilled water, and ultrasonicated for 15 minutes before use to make it evenly suspended.

[0044] BSA formulated at a concentration of 10 -4 mol / L, spare. BSA was purchased from Beijing Suolaibao Biotechnology Company.

[0045] (2) Detection and drawing of standard curve

[0046] After the instrument is preheated stably, take 3mL of pure quantum dots of one of the particle sizes in a cuvette, and perform simultaneous fluorescence detection at 250-750nm, the emission grating and scattering grating are both 5nm, the voltage is 400V, and the scattering intensity is f 0 .

[0047] Then add 10, 20, 30, 40, 50, 60, 70, 80, 90 and 100 μL respectively to a concentration of 10 -4 mol / L BSA solution; each step is tested f...

Embodiment 3

[0049] Example 3 Detection of gamma-globulin

[0050] (1) Preparation of quantum dot solution and protein solution

[0051] The original concentration of quantum dots 1 to 4 is 5×10 -5 mol / L, diluted 60 times with double distilled water, and ultrasonicated for 15 minutes before use to make it evenly suspended.

[0052] The concentration of γ-globulin is 10 -4 mol / L, spare. γ-globulin was purchased from Beijing Suolaibao Biotechnology Company.

[0053] (2) Detection and drawing of standard curve

[0054] After the instrument is preheated stably, take 3mL of pure quantum dots of one of the particle sizes in a cuvette, and perform simultaneous fluorescence detection at 250-750nm, the emission grating and scattering grating are both 5nm, the voltage is 400V, and the scattering intensity is f 0 . Then add 10, 20, 30, 40, 50, 60, 70 and 80 μL of 10 -4 mol / L of γ-globulin, each step is tested for zeroing, and the scattering intensity is F n . It is found that the maximum sc...

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Abstract

The invention discloses a method for quantitatively detecting proteins through quantum dot resonant scattering. The method is a supplement to protein detection methods. The proteins are adsorbed by using quantum dots through an electrostatic effect, resonant scattering light can be generated within a visible light wavelength range to detect standard bovine serum albumins (BSA) and gamma-globulins, a standard curve is drawn by taking the concentration (mol / L) of the proteins as a horizontal coordinate and the highest-intensity change of scattering wavelength as a vertical coordinate, and the concentration of the proteins can be calculated through a regression equation through scattering intensity change. Moreover, the quantum dots with different grain sizes are additionally used for detecting the BSA and the gamma-globulins, good correlation between the concentration of the proteins and the scattering intensity is found, the correlation coefficient reaches 0.95 to 0.99, the gradient reaches 107, the sensitivity is higher, the stability is high, the detection time does not cause a great influence on the result and a new practical method is developed for quantitatively detecting the concentration of biological macromolecular proteins.

Description

technical field [0001] The invention belongs to the technical field of protein detection, and in particular relates to an application method of detecting and analyzing protein content by a quantum dot resonance scattering method. Background technique [0002] Protein is a biomacromolecule composed of amino acids, which is the material basis of life activities. Without protein, there would be no life. Quantitative determination of protein is of great significance for biochemical analysis and clinical medicine. Commonly used protein determination methods include spectrophotometry (Kjeldahl method, biuret method and Coomassie brilliant blue method) and fluorescence photometry, but most of these methods are not sensitive and stable. With the development of proteomics, people need to quantitatively analyze biological samples or purified proteins with lower concentrations, so it is of great significance to find new detection methods to improve the sensitivity of protein determina...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 开国银罗秀芹鞠冠华刘聪肖建波余锡宾
Owner SHANGHAI NORMAL UNIVERSITY
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