Unmarked chemiluminescent immunosensor and immunoassay method thereof

A chemiluminescence immunoassay and immunosensor technology, which is applied in the field of immunology, can solve the problems of lack of label-free chemiluminescence immunosensor immunoassay methods, and achieve the effects of speeding up the overall analysis and detection time, facilitating label-free detection, and simplifying the immunoassay process

Active Publication Date: 2015-02-04
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is a lack of a cheap, fast and convenie...

Method used

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  • Unmarked chemiluminescent immunosensor and immunoassay method thereof
  • Unmarked chemiluminescent immunosensor and immunoassay method thereof
  • Unmarked chemiluminescent immunosensor and immunoassay method thereof

Examples

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Effect test

Embodiment 1

[0035] The invention provides a method for preparing an immunosensor, comprising the following steps:

[0036] (1) Put the glass piece in piranha acid (H 2 SO 4 :H 2 o 2 =7:3, V / V) for 10-12 hours, fully cleaned with distilled water, blown dry in a nitrogen atmosphere, soaked in 1% γ-(2,3 glycidoxy)propyltrimethoxysilane The silanization reaction is carried out in the toluene solution of GPTMS, and placed at room temperature for 10-12 hours; the glass sheet is 2.1 cm×0.4 cm×0.1 cm; the room temperature is 25° C.

[0037] (2) After the silanization reaction, wash with toluene and ethanol successively and blow dry under nitrogen atmosphere;

[0038] (3) After mixing the gold nanoparticle solution and 2.0wt% chitosan solution, ultrasonic to completely disperse, then mix with chemiluminescence probe and antibody solution; surface of a glass slide, and react at room temperature for 0.5-1h, and then place at 4°C for 10-12h; the chemiluminescence probe is horseradish peroxidase ...

Embodiment 2

[0054] The difference between Example 2 and Example 1 is that: the AFP standard samples of different concentrations are measured in this way, and the linear curve of the AFP standard sample is obtained. The chemiluminescent probe is horseradish peroxidase HRP; the antibody solution is an alpha-fetoprotein AFP antibody solution.

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Abstract

The invention discloses an unmarked chemiluminescent immunosensor and an immunoassay method thereof. The immunoassay method comprises the following steps: (1) after fixing the immunosensor on an immune microreactor, injecting antigen samples into a flowing cell at the speed of 0.5 milliliter per minute, and carrying out on-line incubation to generate immune complexes; (2) flushing the immune complexes by virtue of a buffer solution PBST at the speed of 1 milliliter per minute, so as to remove unreacted immune reagents; and (3) introducing a chemiluminescent substrate solution into the immunosensor at the speed of 0.5 milliliter per minute, and recording generated chemiluminescent signals by virtue of a photomultiplier. The immunosensor is produced by jointly fixing a chemiluminescent probe and unmarked antibodies at a high-biocompatibility solid phase interface and is combined with flow injection, so as to construct an unmarked chemiluminescent immunoassay method having the advantages of low cost, rapidness and convenience.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a label-free chemiluminescence immunosensor and an immunoanalysis method thereof. Background technique [0002] Immunoassay technology is based on the specific reaction between antigen and antibody. It is a modern analytical technology with high specificity, high sensitivity and environmental friendliness. It is considered to be an effective method for analyzing complex sample systems. As one of the most useful analysis techniques, flow injection can improve the shortcomings of traditional analysis methods such as bulky, time-consuming and high labor intensity, and has low sample consumption, fewer processing steps, can be reused, good reproducibility, and high analysis speed. Fast and easy to automate, so it has been widely used in many fields. The invention uses immunoassay technology combined with flow injection to automatically detect antigens and markers in human serum, so as to r...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N21/76
CPCG01N21/76G01N33/552
Inventor 杨占军曹越李娟
Owner YANGZHOU UNIV
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