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Near-infrared electro-generated chemiluminescence immunodetection method

A technology of electrochemistry and luminescence immunity, which is applied in the fields of chemiluminescence/bioluminescence, analysis by causing chemical reactions of materials, and measurement devices, etc. It can solve the problem of inability to effectively eliminate the background interference of biological tissues, high ECL excitation potential, and difficult to overcome Interference and other issues, to avoid background light interference of biological tissue, to avoid electrochemical process interference, high sensitivity and accuracy

Active Publication Date: 2015-01-14
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Near-infrared light has the advantages of good tissue penetration, low background interference, and small optical damage. Therefore, near-infrared analysis technology has broad application prospects in the field of life analysis science. It has become far-reaching; Electrochemiluminescence (ECL) analysis does not require an excitation light source, has a high signal-to-noise ratio, and has higher sensitivity than conventional fluorescence analysis. It is an analysis that has been widely used in many research fields technology; but because the electrochemiluminescent radiation of traditional ECL materials is in the visible region, such as luminol and [Ru(bpy) 3 ] 2+ (bpy=2,2′-bipyridyine), etc., so the sensing application of ECL analysis technology has been limited to the visible light region
[0003] With the advancement of nano-fabrication technology, near-infrared quantum dots show great potential in the development of new near-infrared ECL emitters; however, the current research on near-infrared quantum dot ECL is mainly carried out on the cathode, but it is difficult to make breakthroughs on the anode, and its ECL The excitation potential is high and it is difficult to overcome the interference of complex electrochemical reactions and dissolved oxygen in biological tissues in practical applications; at the same time, the near-infrared ECL radiation of quantum dots used in the prior art is concentrated around 700nm, which cannot effectively eliminate the background of biological tissues Interference (Jing Wang, Heyou Han, Xiaochun Jiang, Liang Huang, Lina Chen, and Na Li Anal.Chem., 2012, 84(11), 4893–4899); therefore, seeking to overcome complex electrochemical reactions and biological tissue background Interfering electrochemiluminescence immunoassay methods have become a constant pursuit of the industry

Method used

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Examples

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Embodiment 1

[0045] A near-infrared electrochemiluminescence immunoassay method, the steps are as follows:

[0046] A gold electrode with a diameter of 5.0mm is selected as the carrier material, and alpha-fetoprotein antigen is used as the detection object:

[0047] Step 1. Label the secondary antibody with near-infrared quantum dots as a marker:

[0048] Disperse cadmium telluride (CdTe) quantum dots with carboxyl groups on the surface (the fluorescence radiation wavelength is 780nm) in a solution containing 100 mg / mL 1-ethyl-3-3-dimethylaminopropyl carbodiimide and 100 mg / mL N -In the phosphate buffer of hydroxysuccinimide, add the secondary antibody, vortex to mix and oscillate evenly, react at room temperature, centrifuge the reaction solution, and wash 3 times with phosphate buffer to obtain the quantum dot-labeled secondary antibody;

[0049] Step 2, surface treatment of gold electrodes:

[0050] After polishing the gold electrode with α-alumina polishing powder (0.5-0.7 μm), immer...

Embodiment 2

[0056] A near-infrared electrochemiluminescence immunoassay method, the steps are as follows:

[0057] A gold electrode with a diameter of 5.0mm is selected as the carrier material, and alpha-fetoprotein antigen is used as the detection object:

[0058] Step 3: In the second step, 10 μL of phosphate buffer solution containing 40.0 ng / mL alpha-fetoprotein antigen (Ag) was selected to be drip-coated on the electrode, and other steps were the same as in Example 1.

Embodiment 3

[0060] A near-infrared electrochemiluminescence immunoassay method, the steps are as follows:

[0061] A gold electrode with a diameter of 5.0mm is selected as the carrier material, and alpha-fetoprotein antigen is used as the detection object:

[0062] Step 3: In the second step, 10 μL of phosphate buffer solution containing 10.0 ng / mL alpha-fetoprotein antigen (Ag) or treated patient serum sample was selected to be drip-coated on the electrode, and other steps were the same as in Example 1.

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Abstract

The invention discloses a near-infrared electro-generated chemiluminescence immunodetection method and belongs to the technical field of electro-generated chemiluminescence immunodetection. The method includes: step (1) connecting near-infrared quantum dots and secondary antibodies, (2) performing sandwich immune reaction and manufacturing an electro-generated chemiluminescence immunosensor and (3) using the immunosensor to perform immunodetection. The near-infrared electro-generated chemiluminescence immunodetection method is simple in steps, good in operability and capable of effectively avoiding potential electrochemical process interference and biological tissue background light interference and has high sensitivity and accuracy and good repeatability.

Description

Technical field: [0001] The invention relates to a near-infrared electrochemiluminescence immunodetection method, in particular to an electrochemiluminescence immunodetection method using near-infrared cadmium telluride (CdTe) quantum dots as markers, which belongs to electrochemiluminescence immunodetection technology field. Background technique: [0002] Near-infrared light has the advantages of good tissue penetration, low background interference, and small optical damage. Therefore, near-infrared analysis technology has broad application prospects in the field of life analysis science. It has become far-reaching; Electrochemiluminescence (ECL) analysis does not require an excitation light source, has a high signal-to-noise ratio, and has higher sensitivity than conventional fluorescence analysis. It is an analysis that has been widely used in many research fields technology; but because the electrochemiluminescent radiation of traditional ECL materials is in the visible...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/531G01N21/76
Inventor 邹桂征刘淑风梁国栋
Owner SHANDONG UNIV
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