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Method for coupling acridinium ester with antibody and application of method

An acridinium ester and antibody technology, applied in the field of immune analysis, can solve the problems of specificity to be improved, false positives, etc., and achieve the effect of reducing false positives and improving specificity

Pending Publication Date: 2022-07-01
GUANGDONG FAPON BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, acridinium ester-labeled chemiluminescent detection often has false positive results, and the specificity of the detection needs to be improved

Method used

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  • Method for coupling acridinium ester with antibody and application of method
  • Method for coupling acridinium ester with antibody and application of method
  • Method for coupling acridinium ester with antibody and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A method for an acridine ester conjugated antibody, comprising the following steps:

[0041] (1) Antibody treatment:

[0042] Take a certain amount of the antibody to be labeled, and purify it into carbonate CBS (20mM CB and 150mM NaCl pH9.5) buffer solution by desalting replacement, divide the purified antibody into two equal parts, and take one part of the antibody solution and add EDTA Disodium, use CBS solution to set the antibody concentration to 2mg / mL, and the working concentration of EDTA disodium is 5mM. After the antibody and disodium EDTA are thoroughly mixed, react at room temperature for half an hour; another antibody solution is directly set to CBS solution to The same concentration of 2mg / ml was used for later use.

[0043](2) Conjugation of antibody and acridine ester:

[0044] 2.1. Measure 12 mL of CBS (20 mM CB and 150 mM NaCl pH 9.5) buffer solution into a centrifuge tube;

[0045] 2.2. Add 1 mg of the antibody to be labeled and mix well;

[0046]...

Embodiment 2

[0050] A method for acridinium ester coupling antibody is roughly the same as Example 1, the difference is that the working concentration of disodium EDTA is different, and the difference is as follows:

[0051] The working concentration of disodium EDTA was 4 mM.

Embodiment 3

[0053] A method for acridinium ester coupling antibody is roughly the same as Example 1, the difference is that the working concentration of disodium EDTA is different, and the difference is as follows:

[0054] The working concentration of disodium EDTA was 3 mM.

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Abstract

The invention discloses a method for coupling acridinium ester with an antibody and application of the method, and relates to the technical field of immunoassay, the method comprises the following steps: before the antibody is coupled with the acridinium ester, the antibody and EDTA disodium are subjected to a mixed reaction, compared with the prior art, the antibody treated by the EDTA disodium is coupled with the acridinium ester and is detected, and the detection sensitivity is high. The false positive phenomenon in chemiluminescence immunodetection can be effectively avoided or reduced, and the detection specificity is improved.

Description

Technical field [0001] The present invention relates to the technical field of immunoassay, and specifically to an acridinium ester-coupled antibody method and its application. Background technique [0002] Chemiluminescence immunoassay (CLIA) is a detection and analysis technology that combines highly sensitive chemiluminescence assay technology with highly specific immune reactions and is used for the detection and analysis of various antigens or antibodies. It is the latest immunoassay technology developed after radioimmunoassay, enzyme immunoassay, fluorescence immunoassay and time-resolved fluorescence immunoassay. [0003] Acridinium ester is a type of chemical substance that can be used as a chemiluminescence marker. Under alkaline conditions, acridinium ester molecules are attacked by hydrogen peroxide and can generate dioxyethane. Dioxyethane is unstable and decomposes into CO 2 And N-methylacridone in the electronically excited state emits photons with a wavelengt...

Claims

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Application Information

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IPC IPC(8): G01N33/53
CPCG01N33/53
Inventor 张洁旋
Owner GUANGDONG FAPON BIOTECH CO LTD
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