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Immunoassay reagent kit based on flexible core-shell quantum dot coupling marker and application method of immunoassay reagent kit

A core-shell quantum dot and immunodetection technology, applied in the field of immunodetection, can solve the problems of low sensitivity and poor specificity of immunodetection, and achieve the effects of high sensitivity, high specificity and high fluorescence efficiency.

Inactive Publication Date: 2014-12-17
CHANGCHUN INST OF OPTICS FINE MECHANICS & PHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to solve the technical problems of low sensitivity and poor specificity in the immunodiagnosis kit based on quantum dot labeling in the prior art, and to provide an immunoassay kit based on a core-shell quantum dot flexible coupling marker and its Instructions

Method used

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  • Immunoassay reagent kit based on flexible core-shell quantum dot coupling marker and application method of immunoassay reagent kit
  • Immunoassay reagent kit based on flexible core-shell quantum dot coupling marker and application method of immunoassay reagent kit
  • Immunoassay reagent kit based on flexible core-shell quantum dot coupling marker and application method of immunoassay reagent kit

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preparation example Construction

[0030] The preparation method of the core-shell quantum dots of the present invention belongs to the prior art, and the method utilizes continuous ion layer adsorption reaction technology (SILAR) to realize the core-shell coating of the quantum dots. Adopt the principle of one-pot method, add nuclear raw materials into the same reaction device, obtain bare-nuclear quantum dots with different particle sizes by controlling the time of nuclear growth, then add shell raw materials drop by drop, use SILAR technology for shell coating, and finally The samples were centrifuged and purified to obtain core-shell quantum dots. For specific methods, refer to the article J.Phys.Chem.C2008, 112, 8587-8593. By changing the particle size of the quantum dots, the tuning of the fluorescent color can be realized. It is more preferable to use green light quantum dots with a bare core size of less than 3nm (the fluorescence emission band is at 500-560nm), and the bare core size is 3-4nm (the fluor...

Embodiment 1

[0044] Draw the standard curve:

[0045] (1) Mix 1nmol of CdTe / CdS core-shell quantum dots with a bare core size less than 3nm and 100nmol of EDC condensing agent to activate the carboxyl groups on the surface of the quantum dots, stir at room temperature for 30min, then add 1nmol of protein G, stir at room temperature for 2h, and use NAP -5 column separation and purification to obtain the covalent connection product of quantum dot-protein G;

[0046] (2) Mix 8 pmol of the covalently linked product of quantum dot-protein G with 8 pmol of hepatitis B surface antibody, and stir at room temperature for 1 hour to obtain a flexible coupling labeling product of quantum dot-protein G-HBsAb;

[0047] (3) Take 8 microwell plates coated with 1 pmol hepatitis B surface antibody respectively, add 200 μ L of buffer solution containing HBsAg respectively, the concentration of HBsAg is respectively 0, 0.3, 0.6, 1.25, 2.5, 5, 10, 20 ng / mL Respectively immunoreact at 37°C for 0.5h, after wash...

Embodiment 2

[0050] Immunoassay kit based on core-shell quantum dots flexible coupling label: including 1pmol CdTe / CdS core-shell quantum dots with bare core size less than 3nm, 100pmol EDC condensing agent, 1pmol protein G, 1pmol hepatitis B surface antibody and coating 1pmol microwell plate of hepatitis B surface antibody;

[0051] The method of use of the kit is:

[0052] (1) Mix 1pmol CdTe / CdS core-shell quantum dots with 100pmolEDC condensing agent to activate the carboxyl groups on the surface of the quantum dots, stir at room temperature for 30min, then add 1pmol protein G, stir at room temperature for 2h, separate and purify with NAP-5 column to obtain Quantum dot-protein G covalently linked product;

[0053] (2) Mix 1 pmol of quantum dot-protein G covalently linked product with 1 pmol of hepatitis B surface antibody, and stir at room temperature for 1 hour to obtain a flexible coupling labeling product of quantum dot-protein G-HBsAb;

[0054] (3) Add 200 μL of the test solution ...

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Abstract

The invention discloses an immunoassay reagent kit based on a flexible core-shell quantum dot coupling marker and an application method of the immunoassay reagent kit, belongs to the technical field of immunoassay, and solves the technical problems that a detection kit based on the quantum dot marker and an antibody molecules are biologically marked through the ways such as electrostatic adsorption effect and direct covalent linkage, the sensitivity of the immunoassay is low and the specificity is poor in the prior art. The reagent kit comprises protein, an EDC condensating agent, a core-shell quantum dot, a first antibody of the to-be-detected antigen, and a microporous plate covering a second antibody of the to-be detected antigen, wherein the first antibody of the to-be-detected antigen can be specifically combined with the protein. The reagent kit is high in sensitivity and specificity, simple and sensitive to use, suitable for detecting samples such as blood samples, urine samples and sputum samples and applicable to the fields such as food security, environment monitoring, medical diagnosis, and sanitation epidemic prevention; the detection result is relatively clear.

Description

technical field [0001] The invention belongs to the technical field of immunoassay, and in particular relates to an immunoassay kit based on a core-shell quantum dot flexible coupling marker and a use method thereof. Background technique [0002] At present, enzyme-linked immunoassay kits using enzyme-catalyzed substrates for color development have been widely used. The basic principle is to first coat the corresponding antibody of the antigen in the test object on the microwell plate of the kit, add the sample to be tested, and make the sample After the antigen of the detection object is immunoreacted with the antibody coated on the microwell plate, it is combined with another antibody molecule labeled with the enzyme to form a sandwich structure, and finally the substrate molecule is added to display the substrate through the high-efficiency catalytic performance of the enzyme, so as to determine the detection result. This technique is to add an enzyme-labeled antibody th...

Claims

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Application Information

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IPC IPC(8): G01N33/532G01N21/64
CPCG01N33/533G01N21/64G01N33/5761
Inventor 曾庆辉
Owner CHANGCHUN INST OF OPTICS FINE MECHANICS & PHYSICS CHINESE ACAD OF SCI
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