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Homogeneous immune analysis nano device construction method

A nano-device and immune analysis technology, which is applied in the direction of chemical reaction analysis of materials, material inspection products, chemiluminescence/bioluminescence, etc., can solve the complex structure of the fluorescence resonance energy transfer analysis system, expensive instruments, and increased background Interference and other problems, to achieve the effect of wide luminous range, easy operation, good water solubility and stability

Inactive Publication Date: 2007-10-24
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the structure of the fluorescence resonance energy transfer analysis system is complex and the instrument is expensive
In addition, autofluorescence in organisms will increase background interference, and its application is limited

Method used

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  • Homogeneous immune analysis nano device construction method
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) sodium telluride hydride preparation:

[0032] Put 80 mg of tellurium powder, 80 mg of sodium borohydride and 2 ml of water in a 10 ml small flask and react at room temperature for 8 hours to prepare a sodium telluride hydride solution.

[0033] (2) Synthesis of water-soluble cadmium telluride quantum dots:

[0034]Mix 0.00125 mol / L cadmium chloride and 0.003 mol / L mercaptopropionic acid to form a solution, adjust the pH value to 8.0-9.0 with sodium hydroxide solution, pass nitrogen gas to remove oxygen for 30 minutes, stir vigorously and in nitrogen gas Under protection, inject sodium hydride solution to form cadmium telluride monomer solution. The molar ratio of added divalent cadmium ions: sodium hydride telluride: mercaptopropionic acid is 2:1:4.8. Put the obtained precursor solution into a microwave digestion tank and heat it by microwave radiation. The reaction time is controlled for 0.5-2 hours to obtain the water-soluble cadmium telluride quantum dot with...

Embodiment 2

[0042] (1) The preparation of sodium telluride hydride is the same as in Example 1.

[0043] (2) The synthesis of water-soluble cadmium telluride quantum dots is the same as in Example 1.

[0044] (3) Preparation of cadmium telluride quantum dot-cancer antigen 125 (CA125) biomarker

[0045] Mix 0.3 mg / ml cadmium telluride quantum dots, 0.6 mg / ml EDC and 0.3 mg / ml CA125 in 0.01 mol / L phosphate buffer solution with a pH value of 7.0, and react the mixed solution at room temperature in the dark for 2 -4 hours. Quantum dot biomarkers were purified by ultrafiltration. Finally, store it in the refrigerator at 4 degrees Celsius for later use.

[0046] (4) Preparation of CA125 antibody-peroxidase marker

[0047] Get 0.2 mg / ml peroxidase solution, 0.2 mg / ml EDC solution and 0.3 mg / ml N-hydroxysuccinimide in 0.01 mol / liter phosphate buffer solution with a pH value of 7.0 for about 15 minutes, The reaction solution was purified by ultrafiltration. Then add the CA125 antibody soluti...

Embodiment 3

[0051] (1) The preparation of sodium telluride hydride is the same as in Example 1.

[0052] (2) The synthesis of water-soluble cadmium telluride quantum dots is the same as in Example 1.

[0053] (3) Preparation of cadmium telluride quantum dot-cancer antigen 19-9 (CA19-9) biomarker

[0054] Mix 0.05 mg / ml cadmium telluride quantum dots, 0.1 mg / ml EDC and 0.05 mg / ml CA19-9 in 0.01 mol / L phosphate buffer solution with a pH value of 7.0, and store the mixed solution at room temperature in the dark React for 2-4 hours. Quantum dot biomarkers were purified by ultrafiltration. Finally, store it in the refrigerator at 4 degrees Celsius for later use.

[0055] (4) Preparation of CA19-9 antibody-peroxidase marker

[0056] Containing 0.04 mg / ml peroxidase solution, 0.04 mg / ml EDC solution and 0.06 mg / ml N-hydroxysuccinimide in 0.01 mol / L, pH value of 7.0 phosphate buffer for about After 15 minutes, the reaction solution was purified by ultrafiltration. Then, 0.1 mg / ml CA19-9 ant...

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Abstract

The invention relates to a method for building uniform immunity analyzing nanometer device, composed of a hyperoxide enzyme, an immunity compound, and an energy acceptor nanometer particle cadmium telluride quantum point, which uses the cadmium telluride quantum point as core lighting based on chemical lighting resonance energy transfer theory to be connected with an antigen or an antibody via static or covalence function, and the hyperoxide enzyme is connected with the antibody via covalence, to obtain the biological nanometer device based on antigen-antibody immunity reaction. The invention has better solubility and stability, adjustable emitting wavelength, low background interface, strong light signal via chemical lighting resonance energy transfer, and the application in uniform immunity analysis, without external laser light source.

Description

technical field [0001] The invention relates to a method for constructing a homogeneous immunoassay nano-device. Based on the principle of chemiluminescence resonance energy transfer, the nano-device for homogeneous immunoassay is constructed, and belongs to the technical field of biomolecular nano-device preparation and bioanalysis. Background technique [0002] Immunoassay is a highly selective analytical method based on the specific binding reaction between antibodies and antigens. It is widely used in clinical analysis, food analysis, environmental analysis and other fields. Currently, immunoassays mainly use fluorescence, chemiluminescence and radiation detection methods. Among them, chemiluminescence detection has the characteristics of simple detection equipment and high sensitivity, but its selectivity is poor, and the self-luminescence of the bulk solution interferes greatly. In addition, the current immunoassay mainly adopts the heterogeneous mode, which requires ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N21/76
Inventor 任吉存黄香宜
Owner SHANGHAI JIAO TONG UNIV
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