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Improved synthesis method of heavy metal lead artificial antigen and application of DOTA (2-S-(4-amino-benzene)-1,4,7,10-tetraazacyclononane-1,4,7,10-tetraacetic acid) to preparation of heavy metal lead artificial antigen reagent

A technology of artificial antigen and synthesis method, which is applied in the direction of peptide preparation method, chemical instrument and method, hybrid peptide, etc., can solve the problem of weak characteristic functional groups, weak antigenic determinant antigenic characteristics, affecting high specificity and high Sensitivity, antibody preparation efficiency and other issues, to achieve high affinity and strong specificity

Active Publication Date: 2018-04-24
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These conventional chelating agents are due to the ring-opening and straight-chain structures combined with heavy metal ions, the complex of heavy metal ions and chelating agents as the antigen recognition site is relatively simple in spatial structure, and the characteristic functional group characteristics are not strong, resulting in the prepared The antigenic determinant antigenic characteristics are not strong, which directly affects the preparation efficiency of highly specific and highly sensitive antibodies

Method used

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  • Improved synthesis method of heavy metal lead artificial antigen and application of DOTA (2-S-(4-amino-benzene)-1,4,7,10-tetraazacyclononane-1,4,7,10-tetraacetic acid) to preparation of heavy metal lead artificial antigen reagent
  • Improved synthesis method of heavy metal lead artificial antigen and application of DOTA (2-S-(4-amino-benzene)-1,4,7,10-tetraazacyclononane-1,4,7,10-tetraacetic acid) to preparation of heavy metal lead artificial antigen reagent
  • Improved synthesis method of heavy metal lead artificial antigen and application of DOTA (2-S-(4-amino-benzene)-1,4,7,10-tetraazacyclononane-1,4,7,10-tetraacetic acid) to preparation of heavy metal lead artificial antigen reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Weigh 7 mg DOTA, dissolve it in 2 ml 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) solution (0.01 mol / L, pH7.4), and prepare DOTA chelating agent solution, this reaction solution is A solution;

[0028] Weigh 124.19 mg lead nitrate and dissolve it in 5 ml ultrapure water to prepare 7.5X 10-2 The lead nitrate solution of M, this reaction solution is B liquid;

[0029] Pipette 170 μl of solution B and add it dropwise to solution A, and react in the dark for 3 hours at room temperature, and this reaction solution is solution C;

[0030] Add 630 μl, 20 mM glutaraldehyde solution dropwise to solution C, and react overnight at room temperature in the dark, this reaction solution is solution D;

[0031] Weigh 20 mg of bovine serum albumin (BSA) and dissolve it in 3 ml of HEPES, stir magnetically at room temperature and mix well, this reaction solution is E solution;

[0032] Add solution D to solution E dropwise, and react in the dark for 24 hours at room temperature, ...

Embodiment 2

[0036] Weigh 7 mg DOTA, dissolve it in 2 ml 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) solution (0.01 mol / L, pH7.4), and prepare DOTA chelating agent solution, this reaction solution is A solution;

[0037] Weigh 124.19 mg lead nitrate and dissolve it in 5 ml ultrapure water to prepare 7.5X 10-2 The lead nitrate solution of M, this reaction solution is B liquid;

[0038] Pipette 170 μl of solution B and add it dropwise to solution A, and react in the dark for 3 hours at room temperature, and this reaction solution is solution C;

[0039] Add 630 μl, 20 mM glutaraldehyde solution dropwise to solution C, and react overnight at room temperature in the dark, this reaction solution is solution D;

[0040] Weigh 20 mg of chicken ovalbumin OVA and dissolve in 3 ml of HEPES, stir magnetically at room temperature and mix well, this reaction solution is liquid E;

[0041] Add solution D to solution E dropwise, and react in the dark for 24 hours at room temperature, then add 170...

Embodiment 3

[0045] Weigh 7 mg DOTA, dissolve it in 2 ml 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) solution (0.01 mol / L, pH7.4), and prepare DOTA chelating agent solution, this reaction solution is A solution;

[0046] Weigh 124.19 mg lead nitrate and dissolve it in 5 ml ultrapure water to prepare 7.5X 10-2 The lead nitrate solution of M, this reaction solution is B liquid;

[0047] Pipette 170 μl of solution B and add it dropwise to solution A, and react in the dark for 3 hours at room temperature, and this reaction solution is solution C;

[0048] Add 630 μl, 20 mM glutaraldehyde solution dropwise to solution C, and react overnight at room temperature in the dark, this reaction solution is solution D;

[0049] Weigh 20 mg of bovine serum albumin (BSA) and dissolve it in 3 ml of HEPES, stir magnetically at room temperature and mix well, this reaction solution is E solution;

[0050] Add liquid D dropwise to liquid E, and react in the dark for 24 hours at room temperature to ob...

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Abstract

The invention provides a synthesis method for preparing a heavy metal lead artificial antigen based on a novel difunctional chelating agent DOTA (2-S-(4-amino-benzene)-1,4,7,10-tetraazacyclononane-1,4,7,10-tetraacetic acid). According to the preparation method, DOTA is taken as the chelating agent, a lead ion chelating agent compound is coupled with BSA (bovine serum albumin) as carrier protein orKHL (human serum albumin), and the artificial antigen is prepared. According to the method, the step of sodium borohydride reduction is added on the original traditional basis, so that the coupling efficiency is improved. The invention further provides an application of DOTA to preparation of the heavy metal lead artificial antigen.

Description

technical field [0001] The invention relates to a synthesis technology of an artificial antigen targeting heavy metal lead using a novel bifunctional chelating agent DOTA as a linking carrier, which belongs to the technical field of artificial antigen preparation in immunochemical technology and can be used for subsequent high-specificity anti-heavy metal lead antibody production Research on preparation and immunoassay techniques, and used in the detection of trace and constant heavy metal lead in environmental samples. Background technique [0002] Heavy metal pollution mainly refers to copper, cadmium, mercury, nickel, chromium, arsenic, zinc, lead and other pollutants. Heavy metals are widely distributed and difficult to degrade. They can enter the human body through the atmosphere, water, and food chain, and interact with proteins and various enzymes in the body to deactivate them and accumulate in certain organs. If they exceed the tolerance limit of the human body , c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K14/77C07K1/107
CPCC07K14/765C07K14/77C07K19/00
Inventor 金仁耀郭建军
Owner ZHEJIANG GONGSHANG UNIVERSITY
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