Application of metallic ion specific response based fluorescence detection method in immune detection

A metal ion and fluorescence detection technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of no application, low sensitivity, application limitations, etc., and achieve the effects of reducing detection costs, improving sensitivity, and improving stability.

Inactive Publication Date: 2015-12-23
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the color development mechanism of this detection technology itself, its sensitivity is low, and it is not suitable for detecting low-concentration analytes,

Method used

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  • Application of metallic ion specific response based fluorescence detection method in immune detection
  • Application of metallic ion specific response based fluorescence detection method in immune detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Using the microtiter plate as the carrier of antigen and antibody, the application of the fluorescence detection method based on the specific response of silver ions in the detection of alpha-fetoprotein (AFP) (see figure 1 ), in the application, the microplate 101 can use microplates with different numbers of holes such as 96-well microplate, 48-well microplate or 384-well microplate, and the microplate contains detachable and non-detachable two There are three models, and the color of the microtiter plate includes black, white and colorless transparent color. The coating antibody 2 used is a monoclonal AFP antibody; the antigen 3 used is an AFP antigen; the silver-labeled antibody used is a polyclonal AFP-labeled antibody 4 labeled with silver nanoparticles 5; the metal ion fluorescent molecule detection solution used is a fluorescent molecule containing silver ions 6 Fluorescence detection solution.

[0026] First, the coated antibody 2 is adsorbed on the microtiter...

Embodiment 2

[0062] Using the microtiter plate as the carrier of antigen and antibody, the application of the fluorescence detection method based on the specific response of metal ions in the detection of serum samples of liver cancer patients and non-liver cancer patients (refer to figure 1 ), the content of each step is basically the same as in Example 1. The difference between embodiment 2 and embodiment 1 is:

[0063] The enzyme label plate 1 adopts a 96-well transparent detachable enzyme label plate, and the antigen 3 used uses serum samples of liver cancer patients and non-liver cancer patients.

[0064] Step (4) Take the serum samples of liver cancer patients and non-liver cancer patients as an example for immunoassay

[0065] The contents of step (4) ①, ②, ④, ⑤ are the same as in Example 1.

[0066] Step (4) ③ Wash the microtiter plate 101 of step (2) three to five times with phosphate buffered saline Tween (PBST), and add 100 μL serum samples of liver cancer patients and non-liv...

Embodiment 3

[0072] The application of the fluorescence detection method based on the specific response of metal ions in the detection of alpha-fetoprotein (AFP) (see figure 2 ), in application, the magnetic beads 102 are used as carriers of antigens and antibodies. The magnetic beads 102 include amino magnetic beads, carboxyl magnetic beads, epoxy-based magnetic beads and streptavidin magnetic beads with a diameter of 50-500nm, the coated antibody 2 used is a monoclonal AFP antibody, and the antigen 3 used is AFP For the antigen, the silver-labeled antibody used is polyclonal AFP antibody 4 labeled with silver nanoparticles 5 , and the metal ion fluorescent molecule detection solution used is a fluorescent detection solution containing silver ion fluorescent molecule 6 .

[0073] First, the coated antibody 2 is linked to the magnetic beads 102 through a chemical reaction.

[0074] Then, the magnetic beads 102 coated with the antibody 2 are combined with the antigen 3 , and further linke...

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Abstract

The invention discloses an application of a metallic ion specific response based fluorescence detection method in immune detection. According to the invention, a traditional metallic ion fluorescence detection method and a metal nanoparticle-label immune technology are combined and applied to the immune detection, the problem of difficulty in connection of traditional fluorescent molecules and biological protein molecules is solved, detection signals are effectively amplified, and the problems caused by strict requirements of the enzyme-catalyzed reaction for experimental conditions are solved; magnetic beads are taken as carriers of antigens and antibodies in the immune detection, rapid separation of to-be-detected samples can be realized, the detection process is simpler, more convenient and more rapid, and accuracy and sensitivity of the detection are improved greatly; a traditional enzyme catalysis process is replaced with the chemical reaction, the fluorescence detection method is introduced into the biological immune detection, so that the reaction time is shortened, the system stability is improved, the detection cost is reduced, and the application can play an important role in immune analysis and detection and clinical research.

Description

technical field [0001] The invention relates to the technical field of biological immune detection, in particular to the application of a fluorescence detection method based on the specific response of metal ions in immune detection. Background technique [0002] The invention utilizes the specific response of fluorescent molecules of various metal ions to metal ions and the specific reaction between fluorescent molecules and metal ions to cause changes in autofluorescence intensity, and applies the changes to immune detection technology. Since the changes in fluorescence intensity caused by different concentrations of metal ions are different, measuring the relative fluorescence intensity with a fluorescence spectrometer can realize quantitative detection in immunoassay. [0003] Fluorescent molecules, because of their high sensitivity up to single-molecule detection, and visible sub-nanometer spatial resolution and sub-millisecond time resolution for sub-particles, can be ...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N33/543
CPCG01N33/533G01N33/54326
Inventor 龙亿涛赵立军马巍韩焕兴于汝佳
Owner EAST CHINA UNIV OF SCI & TECH
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