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Antibody directional marking colloidal gold immune probe, preparation method thereof and lead ion test paper based on antibody directional marking

A technology of colloidal gold and lead ions, which is applied in antibody-directed labeling of colloidal gold immunoprobes and its preparation, and in the field of lead-ion test paper based on antibody-directed labeling, can solve the problems of uncontrollable antibody spatial orientation and low effective labeling of antibodies, and achieve Reduce detection cost, sensitive detection, low detection cost effect

Pending Publication Date: 2022-07-22
WUHAN POLYTECHNIC UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The main purpose of the present invention is to propose a colloidal gold immunoprobe for antibody-directed labeling and its preparation method and lead ion test paper based on antibody-directed labeling, aiming to solve the problem that the existing lead ion test paper cannot control the spatial orientation of antibodies and the effective labeling of antibodies is low. question

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  • Antibody directional marking colloidal gold immune probe, preparation method thereof and lead ion test paper based on antibody directional marking
  • Antibody directional marking colloidal gold immune probe, preparation method thereof and lead ion test paper based on antibody directional marking
  • Antibody directional marking colloidal gold immune probe, preparation method thereof and lead ion test paper based on antibody directional marking

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preparation example Construction

[0052] Also, see figure 2 , the present invention provides a method for preparing an antibody-directed labeled colloidal gold immunoprobe, and the antibody-directed labeled colloidal gold immunoprobe 100 prepared by the method for preparing an antibody-directed labeled colloidal gold immunoprobe is subjected to lead ion concentration The detection can ensure the full exposure of the antibody antigen binding site (Fab), improve the effective labeling of the antibody on the surface of colloidal gold, reduce the amount of the antibody, and reduce the detection cost, and the preparation method of the antibody-directed labeled colloidal gold immunoprobe Include the following steps:

[0053] Step S10, adjusting the pH of the colloidal gold particles 1 to 7-10, adding connexin 2 and stirring evenly, then adding bovine serum albumin 4 solution, and centrifuging to obtain a first precipitate.

[0054] In this embodiment, before preparing the first precipitation, the pH value of the c...

Embodiment 1

[0073] (1) Preparation of antibody-directed labeled colloidal gold immunoprobes

[0074] Add potassium carbonate solution dropwise to the colloidal gold particles, adjust the pH to 7.3, add recombinant protein G according to the mass ratio of colloidal gold particles and recombinant protein G to 100:18, react at 37°C for 30 min, and then add The bovine serum albumin solution with a concentration of 0.5% was reacted at 37° C. for 30 min, and then centrifuged at 9500 r / min for 13 min. After 7 times of centrifugation, the first precipitate was obtained.

[0075] The first precipitate was dissolved in a phosphate buffer with a concentration of 0.01 mol / L to obtain a colloidal gold-connexin complex.

[0076] According to the mass ratio of colloidal gold particles and anti-lead ion monoclonal antibody to 100:17.5, anti-lead ion monoclonal antibody was added to the colloidal gold-connexin complex, and the reaction was carried out at 37 °C for 120 min, and then at 8000 r / Centrifuge ...

Embodiment 2

[0085] (1) Preparation of antibody-directed labeled colloidal gold immunoprobes

[0086] Add potassium carbonate solution dropwise to the colloidal gold particles, adjust the pH value to 9.8, add recombinant protein A according to the mass ratio of colloidal gold particles and recombinant protein A to 100:3, react at 28 °C for 50 min, and then add The bovine serum albumin solution with a concentration of 0.55% was reacted at 28° C. for 50 min, and then centrifuged at 11,000 r / min for 12 min, and the first precipitate was obtained after centrifugation for 6 times.

[0087] The first precipitate was dissolved in a phosphate buffer with a concentration of 0.01 mol / L to obtain a colloidal gold-connexin complex.

[0088] According to the mass ratio of colloidal gold particles and anti-lead ion monoclonal antibody is 100:12.3, add anti-lead ion monoclonal antibody to the colloidal gold-connexin complex, react at 28°C for 150min, and then at 10000r / min Centrifuged for 8 min at 100 r...

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Abstract

The invention discloses an antibody directional labeling colloidal gold immune probe, a preparation method thereof and lead ion test paper based on antibody directional labeling. The antibody directional labeling colloidal gold immune probe comprises the following components: colloidal gold particles, connexin and an anti-lead ion monoclonal antibody, wherein the connexin is used for connecting the colloidal gold particles and the anti-lead ion monoclonal antibody, and the connexin comprises a protein for specifically recognizing an Fc fragment of the monoclonal antibody. The invention aims to solve the problems that the existing lead ion test paper cannot control the spatial orientation of an antibody and the effective label of the antibody is low.

Description

technical field [0001] The invention relates to the technical field of lead ion testing, in particular to an antibody-directed labeled colloidal gold immunoprobe, a preparation method thereof, and a lead ion test paper based on the antibody-directed labeling. Background technique [0002] Lead is one of the heavy metal elements that has a huge toxic effect on human health. It is widely used in paint pigments, batteries and building materials. It can accumulate in the environment and enter the human body through the food chain, seriously endangering people's health. Therefore, the detection technology research of heavy metal lead is of great significance to protect people's health. [0003] At present, the main detection techniques of heavy metal lead include atomic absorption spectrometry, atomic emission spectrometry, inductively coupled plasma mass spectrometry, atomic fluorescence spectrometry, stripping voltammetry and dithizone colorimetry. The above detection method ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/531G01N33/558G01N33/577G01N33/58
CPCG01N33/5308G01N33/531G01N33/577G01N33/585G01N33/558
Inventor 许琳李杨阳曲文莉杨庆王桥柳鑫宫智勇
Owner WUHAN POLYTECHNIC UNIVERSITY
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