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Monoclonal antibody capable of resisting heavy metal copper and preparation method thereof

A monoclonal antibody and antigen technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, chemical instruments and methods, etc., can solve the problems of cumbersome detection steps, expensive detection instruments, high detection costs, etc., to ensure food safety, Sensitive, specific, high titer effect

Inactive Publication Date: 2011-05-25
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional heavy metal detection methods mostly use chemical instruments, such as AAS, ICP-AES or electrochemical methods, etc. The detection instruments are expensive, and the samples have to be digested by wet method or microwave digestion to measure the concentration of single heavy metals one by one. Although the measurement accuracy can reach mg / kg or higher, but the detection steps are cumbersome, the detection cost is high, and it takes about 2 days. It does not meet the needs of quick and easy in practice, and it is difficult to adapt to the environment and on-site random inspection of market products and the requirements of fast customs clearance for import and export of products.

Method used

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  • Monoclonal antibody capable of resisting heavy metal copper and preparation method thereof
  • Monoclonal antibody capable of resisting heavy metal copper and preparation method thereof
  • Monoclonal antibody capable of resisting heavy metal copper and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1, the preparation of antigen and incomplete coating former

[0053] 1. Preparation

[0054] p-Scn-Bn-DTPA was purchased from Macrocyclics in the United States, the product number is B-305.

[0055] Product name: p-SCN-Bn-DTPA

[0056] English name: 2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid

[0057] Alias: S-2-(4-Isothiocyanatobenzyl)-Diethylenetriamine Pentaacetic Acid

[0058] Molecular formula: C 22 h 28 N 4 o 10 S.3HCl.

[0059] HEPES buffer solution with a pH value of 9.73: 1.4165g HEPES, dissolved in deionized water, adjusted to pH 9.73 with 10mol / L KOH, adjusted to 50mL, filtered and sterilized after preparation, and stored at 4°C. (The concentration is 1.4165g / 50mL).

[0060] HEPES buffer solution with a pH value of 7.4: 14.165g HEPES, dissolved in deionized water, adjust the pH to 7.4, and dilute to 500mL. (The concentration is 1.4165g / 50mL).

[0061] Solution A: 11 g of copper flakes were dissolved in 100 mL of 68% by ...

Embodiment 2

[0111] Example 2, Preparation and Functional Verification of Monoclonal Antibody

[0112] 1. Preparation of monoclonal antibodies

[0113] (1) Preparation of hybridoma cell line C9D11

[0114] 1. Immunization of mice and selection of fusion mice

[0115] 1) Basic immunization: For the first immunization, select 8-week-old BABL / c mice with a body weight of about 23-25 ​​g) to take Cu-DTPA-KLH with a protein concentration of 6.314 mg / mL and an equal volume of Freund's complete adjuvant The agent was thoroughly mixed and emulsified for 1 to 2 hours until it was dripped into water and did not diffuse. Each BABL / c was intraperitoneally injected with 200 μg of Cu-DTPA-KLH antigen. Three weeks later, the second immunization was carried out, and the same dose of antigen was fully emulsified with the same volume of Freund's incomplete adjuvant, and then immunization was performed every two weeks, and the method was the same as the second immunization. After the third immunization an...

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PUM

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Abstract

The invention discloses a monoclonal antibody capable of resisting heavy metal copper. The monoclonal antibody is secreted by the hybridoma cell strain C9D11 with the CCTCC NO of C201087. The monoclonal antibody capable of resisting heavy metal copper has high potency. The antibody of the invention can be used for the immunological test of the residual copper ions. The advantages of the antibody, namely high speed, low price, sensitivity and high specificity can be utilized to develop a convenient field testing method, thus the method is suitable for the sampling test of agricultural and livestock products and the rapid test of customs clearance. The monoclonal antibody has practical significance in increasing the efficiency and quality of risk assessment and ensuring the food safety.

Description

technical field [0001] The invention relates to a monoclonal antibody against heavy metal copper and a preparation method thereof. Background technique [0002] With the advancement of science and technology and the development of industry, people's living standards have been greatly improved. However, with the rapid development of industry, it has also brought a series of environmental problems. The Itai disease that occurred in the Jinzugawa River Basin in Toyama Prefecture, Japan from 1931 to 1972, and the Minamata disease that occurred in the fishing village of Mizuo Bay in Kumamoto Prefecture, Japan in 1953 were both caused by heavy metal contamination of food. heavy metal pollution concerns. [0003] What are heavy metals? Heavy metals refer to about 60 elements with a density above 4.0 or 45 elements with a density above 5.0. The heavy metals referred to in terms of environmental pollution mainly refer to mercury, cadmium, lead, chromium and metalloid arsenic with...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44C07K14/765C07K14/795G01N33/577C12R1/91
Inventor 赵丽李霞杨慧
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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