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Preparation method of signal amplifying type quantum dot immune fluorescent probe and application of signal amplifying quantum dot immune fluorescent probe

A technology of immunofluorescence and quantum dots, which is applied in the field of immunoassay, can solve the problems of low detection sensitivity and low signal, and achieve the effect of high detection sensitivity, good repeatability and uniform particle size

Inactive Publication Date: 2013-10-23
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the problems of low signal and low detection sensitivity of current quantum dot immunofluorescence probes, and to provide a method for preparing signal-amplified quantum dot immunofluorescence probes. Another purpose of the present invention is to provide the method for preparing Application of Quantum Dot Immunofluorescence Probe in Fluorescence Immunoassay (Kit)

Method used

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Embodiment 1

[0052] Embodiment 1: Preparation of quantum dot-polymer composite nanosphere

[0053] The preparation of CdSe / ZnS quantum dot-polymer composite nanosphere comprises the following steps:

[0054] A. Oil phase components: CdSe / ZnS quantum dots and polystyrene-maleic anhydride copolymer are dispersed in chloroform solution, the concentration of quantum dots is 0.1μmol / L, and the concentration of polymer is 0.05mg / mL;

[0055] B. Water phase components: Polyvinyl alcohol and sodium dodecylsulfonate are configured into an aqueous solution, wherein the concentration of polyvinyl alcohol is 2.5%, and the concentration of sodium dodecylsulfonate is 0.5%;

[0056] C. In a water bath at 4°C, under magnetic stirring, add the oil phase solution to the water phase solution, and keep stirring for 60 minutes, and then use a sonicator to sonicate the solution for 10 minutes to obtain a uniform and stable microemulsion.

[0057] D, at room temperature, the microemulsion is placed in an open c...

Embodiment 2

[0063] Embodiment 2: Preparation of quantum dot-polymer composite nanosphere

[0064] A. Oil phase components: CdSe quantum dots and polystyrene-acrylic acid copolymer are dispersed in chloroform solution, the concentration of quantum dots is 2.5 μmol / L, and the concentration of polymer is 1.0 mg / mL;

[0065] B. Water phase components: Polyvinyl alcohol and sodium dodecylsulfonate are configured into an aqueous solution, wherein the concentration of polyvinyl alcohol is 0.5%, and the concentration of sodium dodecylsulfonate is 0.25%;

[0066] C. In a water bath at 4°C, under magnetic stirring, add the oil phase solution to the water phase solution, continue stirring for 10 minutes, and then use a sonicator to sonicate the solution for 3 minutes to obtain a uniform and stable microemulsion.

[0067] D, at room temperature, the microemulsion is placed in an open container, under magnetic stirring, the solvent is gradually volatilized, the volatilization time is 24 hours, centrif...

Embodiment 3

[0072] Embodiment 3: Preparation of quantum dot-polymer composite nanosphere

[0073] A. Oil phase components: Disperse CuInS quantum dots and polymethyl methacrylate-methacrylic acid copolymer in chloroform solution, the concentration of quantum dots is 0.1 μmol / L, and the concentration of polymer is 0.05 mg / mL;

[0074] B. Water phase components: Polyvinyl alcohol and sodium dodecylsulfonate are configured into an aqueous solution, wherein the concentration of polyvinyl alcohol is 2.5%, and the concentration of sodium dodecylsulfonate is 0.5%;

[0075] C. In a water bath at 4°C, under magnetic stirring, add the oil phase solution to the water phase solution, and keep stirring for 60 minutes, and then use a sonicator to sonicate the solution for 10 minutes to obtain a uniform and stable microemulsion.

[0076] D, at room temperature, the microemulsion is placed in an open container, under magnetic stirring, the solvent is gradually volatilized, the volatilization time is 6 ho...

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Abstract

The invention belongs to technical field of immunoassay, and particularly relates to a preparation method of a signal amplifying type quantum dot immune fluorescent probe and an application of the signal amplifying type quantum dot immune fluorescent probe. At first, According to the method, a microemulsion solvent evaporation method is adopted for preparing a quantum dot-polymer composite nanosphere with controllable grain size and good dispersibility by utilizing quantum dot and polymer solution; and an antibody is covalently coupled on the surface of the composite nanosphere to obtain the signal amplifying type quantum dot immune fluorescent probe. The invention provides an application of the quantum dot immune fluorescent probe in a fluorescence immunoassay method or a fluorescence immunoassay detection kit. Compared with traditional quantum dot immune fluorescent probe, the quantum dot immune fluorescent probe prepared by the method provided by the invention has stronger signals and good detection sensitivity.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and in particular relates to a preparation method and application of a signal-amplifying quantum dot immunofluorescence probe. Background technique [0002] Immunoassay technology is a detection method based on the principle of specific reaction between antibodies and antigens for qualitative and quantitative analysis of the test object. The immunoassay of low-concentration antigens is still an important problem in the field of clinical diagnosis and inspection and quarantine. The accuracy and accuracy of low-concentration antigens Reliable detection can provide an important scientific basis for early diagnosis and timely treatment of diseases, and one of the methods to improve the limit of immune detection is to prepare highly sensitive immune probes. Fluorescent immunoassay technology uses fluorescent substances to label antibodies or antigen molecules, and detects the intensity changes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/88C09K11/62G01N33/577G01N33/545G01N33/544
Inventor 韩焕兴张鹏飞叶伟民
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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