Lead ion antigen and preparation method and application thereof

A lead ion and antigen technology, used in chemical instruments and methods, animal/human proteins, serum albumin, etc., can solve the problems of expensive detection of lead ions, inability to detect quickly, and high detection costs, and achieve concise and minimal synthesis steps. The effect of low detection limit and low cost of synthesis

Inactive Publication Date: 2011-01-05
CHINA AGRI UNIV
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  • Description
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Problems solved by technology

However, the detection of lead ions by these methods requires expensive instruments and equ

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  • Lead ion antigen and preparation method and application thereof
  • Lead ion antigen and preparation method and application thereof
  • Lead ion antigen and preparation method and application thereof

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Embodiment 1

[0035] Embodiment 1, lead ion antigen and preparation thereof

[0036] figure 1 It is a synthetic route map of the lead ion antigen, and the specific steps are as follows:

[0037] 1. Preparation of lead-p-aminobenzylethylenediaminetetraacetic acid-ovalbumin antigen (Pb-p-aminobenzyl-EDTA-OVA)

[0038] 1. Diazotization: Dissolve 2.5mg of p-aminobenzylethylenediaminetetraacetic acid (Aminobenzyl-EDTA) in 2mL of 1M HCl aqueous solution to obtain solution I; 50ul 0.1% (mass percentage) NaNO 2 aqueous solution, stirred and reacted for 15 minutes to obtain an acidic solution of diazotized p-aminobenzylethylenediaminetetraacetic acid. In the reaction system, p-aminobenzylethylenediaminetetraacetic acid, HCl and NaNO 2 The feeding ratio is 2.5mg: 2.5×10 -4 mol: 0.7×10 -5 mol.

[0039] The structural formula of diazotized p-aminobenzylethylenediaminetetraacetic acid is as shown in formula I:

[0040]

[0041] (Formula I).

[0042] 2. Coupling: The acidic solution of the di...

Embodiment 2

[0067] Embodiment 2, the application of lead-p-aminobenzylethylenediaminetetraacetic acid-ovalbumin antigen (Pb-p-aminobenzyl EDTA-OVA)

[0068] 1. Preparation of antibody using lead-p-aminobenzylethylenediaminetetraacetic acid-ovalbumin antigen

[0069] (1) Take 8-10 week old Bal b / c mice as experimental animals.

[0070] (2) Basic immunization: the Pb-p-aminobenzyl EDTA-OVA antigen solution with a concentration of 1 mg / mL obtained in Example 1 was filtered through a sterile filter and then added with an equal volume of Freund's complete adjuvant, and the mixture was mixed with a magnetic stirrer. Stir well to emulsify until dripping into water does not spread. Bal b / c mice were injected subcutaneously with emulsified complete antigen at multiple points in the abdominal cavity and back, and the injection dose was 0.1 mg emulsified antigen / mouse.

[0071] (3) Booster immunization: 2 weeks after the basic immunization, take 1 mL of Pb-p-aminobenzyl EDTA-OVA antigen solution w...

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Abstract

The invention discloses a lead ion antigen and a preparation method and application thereof. The preparation method of the lead ion antigen comprises the following steps: 1) dissolving chelating agent to the aqueous solution of acid, and adding the aqueous solution of nitrite to carry out diazotization to obtain diazotized chelating agent; 2) mixing the diazotized chelating agent obtained in step 1) with carrier protein for coupling reaction to obtain the conjugate of the chelating agent and the carrier protein; 3) mixing the conjugate of the chelating agent and the carrier protein, which is obtained in step 2) with lead ion solution for complexation, to obtain a complex formed by the lead ion and the conjugate of the chelating agent and the carrier protein. The preparation method of the lead ion antigen and the lead ion antigen obtained with the method have wide prospect in the quick immunological detection application of the lead ion.

Description

technical field [0001] The invention relates to a lead ion antigen and its preparation method and application. Background technique [0002] Heavy metals are important pollutants in the environment and agricultural products. They can be enriched in animals and humans through the food chain, causing serious harm to human health. Lead is one of the more toxic heavy metal pollution, and once it enters the human body, it is difficult to get rid of it. Direct damage to human brain cells, especially the neural plate of the fetus, can cause congenital brain sulcus shallow and mental retardation; it can cause dementia and brain death to the elderly. [0003] At present, the analysis methods of heavy metal lead mainly include atomic absorption spectrometry, inductively coupled plasma emission / mass spectrometry, X-ray fluorescence spectrometry, potential stripping analysis and biosensor detection developed in recent years. However, the detection of lead ions by these methods require...

Claims

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Application Information

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IPC IPC(8): C07K14/765C07K14/77C07K16/44G01N33/566
Inventor 王保民曹振赵洪伟南铁贵高巍谭桂玉王敏孙硕李召虎
Owner CHINA AGRI UNIV
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