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Nanometer silver probe modified by fluorescent molecule, kit thereof, and application thereof in detecting human IgE

A fluorescent molecule and nano-silver technology, which is applied in fluorescence/phosphorescence, luminescent materials, material excitation analysis, etc., can solve the problems of easy expiration of radioisotopes, difficult to obtain double antibodies, and pollution of the environment, so as to achieve good fluorescence enhancement effect and improve detection. Good performance and specificity

Active Publication Date: 2012-12-19
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Commonly used methods for detecting human IgE include radioimmunoassay and enzyme-linked immunoassay. The disadvantages of radioimmunoassay are high cost, long time, easy expiration of radioisotopes and pollution of the environment, and the disadvantage of enzyme-linked immunoassay is the need to use double antibodies. The cost is high, and the double antibody is not easy to obtain

Method used

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  • Nanometer silver probe modified by fluorescent molecule, kit thereof, and application thereof in detecting human IgE
  • Nanometer silver probe modified by fluorescent molecule, kit thereof, and application thereof in detecting human IgE
  • Nanometer silver probe modified by fluorescent molecule, kit thereof, and application thereof in detecting human IgE

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Preparation of nano-silver probes.

[0051] (1) Under the condition of ice bath, add 2mmol / L silver nitrate dropwise to 3mmol / L sodium borohydride solution at a certain speed (the molar ratio between the two is 1:3), and keep stirring until the reaction is complete After the reaction is completed, 7% molar sodium borohydride solution is added and heated to obtain a yellow solution, and the concentration of nano-silver is 2.6nM.

[0052] (2) Add fluorescent molecular chains and aptamer chains in a molar ratio of 3:1 to the nano-silver solution obtained in step (1), and let stand for 18 hours. The ratio of the total molar weight of fluorescent molecular chains and aptamer chains to the molar weight of nano silver is 288.

[0053] The nucleotide sequence of the fluorescent molecular chain is: 5'AAAAAAAAAAAAAAAAAAAAAAAAAA, its 5' end is modified by thiol, and its 3' end is modified by Cy5;

[0054] The nucleotide sequence of the aptamer chain is: 5' AAAAA AAAAA...

Embodiment 2

[0059] Example 2: Preparation of nano-silver probes.

[0060] (1) Under the condition of ice bath, add 20mmol / L silver nitrate dropwise to 30mmol / L sodium borohydride solution at a certain speed (the molar ratio between the two is 1:3), and keep stirring until the reaction is complete After the reaction is completed, 7% molar sodium borohydride solution is added and heated to obtain a yellow solution, and the concentration of nano-silver is 2.6nmol / L.

[0061] (2) Add fluorescent molecular chains and aptamer chains in a molar ratio of 1:1 to the nano-silver solution obtained in step (1), and let stand for 10 hours. The ratio of the total molar weight of fluorescent molecular chains and aptamer chains to the molar weight of nano silver is 100.

[0062] The nucleotide sequence of the fluorescent molecular chain is: 5'AAAAAAAAAAAAAAAAAAAAAAAAAA, its 5' end is modified by thiol, and its 3' end is modified by Cy5;

[0063] The nucleotide sequence of the aptamer chain is: 5' AAAAA...

Embodiment 3

[0068] Example 3: Preparation of nano-silver probes.

[0069] (1) Under the condition of ice bath, add 10mmol / L silver nitrate dropwise to 20mmol / L sodium borohydride solution at a certain speed (the molar ratio between the two is 1:3), and keep stirring until the reaction is complete After the reaction is completed, 7% molar sodium borohydride solution is added and heated to obtain a yellow solution, and the concentration of nano-silver is 2.6nmol / L.

[0070] (2) Add fluorescent molecular chains and aptamer chains in a molar ratio of 99:9 to the nano-silver solution obtained in step (1), and let stand for 24 hours. The ratio of the total molar weight of fluorescent molecular chains and aptamer chains to the molar weight of nano silver is 1000.

[0071] The nucleotide sequence of the fluorescent molecular chain is: 5'AAAAAAAAAAAAAAAAAAAAAAAAAA, its 5' end is modified by thiol, and its 3' end is modified by Cy5;

[0072] The nucleotide sequence of the aptamer chain is: 5' AAA...

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Abstract

The invention discloses a nanometer silver probe modified by fluorescent molecule. The nanometer silver probe uses a nanometer silver ball having a diameter of 10 to 30 nm as a core, and the external surface of the silver ball is bonded with a fluorescent molecule chain and an aptamer chain. The invention further discloses a kit containing the probe and an application in detecting human IgE. The nanometer silver probe provided by the invention can greatly improve sensitivity of fluorescence detection by coupling a fluorescence enhancing liquid, and a linear range of human IgE detection is from 0.5 ng / ml to 10 [mu]g / ml, thereby being low in detection limit and good in specificity. Such nanometer silver probe is mature in synthesis and modification method of, the fluorescence enhancing liquid is cheap and easily available, and sensitivity of human IgE detection can be greatly enhanced by combining the aforementioned two factors. Such signal amplified method provides an excellent method for fluorescence analysis and detection of protein markers.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a nano-silver probe modified by fluorescent molecules, a reagent kit thereof and an application in detecting human IgE. Background technique [0002] Human immunoglobulin IgE is a low-abundance protein that is closely related to human allergic reactions. Monitoring the changes of IgE in the human body is of great significance for the prevention and diagnosis of diseases. Commonly used methods for detecting human IgE include radioimmunoassay and enzyme-linked immunoassay. The disadvantages of radioimmunoassay are high cost, long time, easy expiration of radioisotopes and pollution of the environment, and the disadvantage of enzyme-linked immunoassay is the need to use double antibodies. The cost is high, and the double antibody is not easy to obtain. Aptamers are DNA or RNA molecules with a three-dimensional structure that can specifically recognize targe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C09K11/06
Inventor 许丹科李慧魏霞羌维兵李钟卉
Owner NANJING UNIV
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