Heavy metal enzyme label and application thereof

A technology of heavy metals and enzyme labels, applied in the direction of microorganisms, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of unavailable rapid detection, high detection cost, and high cost, and achieve HRP activity loss, low synthesis cost, The effect of simple preparation method

Inactive Publication Date: 2011-01-19
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the detection of copper ions by these methods requires expensive equipment, high detection costs, time-consuming, and cannot be used for rapid on-site detection.

Method used

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  • Heavy metal enzyme label and application thereof
  • Heavy metal enzyme label and application thereof
  • Heavy metal enzyme label and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1. A kit for detecting whether the sample contains heavy metal copper

[0057] 1. Composition of the kit

[0058] (1) Composition of the kit of the present invention

[0059] 1. Original coating: anti-copper monoclonal antibody, which is the monoclonal antibody secreted by the hybridoma cell line Cu-EDTA 6A9 with deposit number CGMCC No.3987;

[0060] 2. Enzyme-labeled substance: The enzyme-labeled heavy metal copper is a cross-linking product formed by a complex and horseradish peroxidase through covalent bonds; the complex is a combination of p-aminobenzylethylenediaminetetraacetic acid and BSA. The conjugate formed by covalent bond is connected with the heavy metal copper through coordination bond to form a complex.

[0061] 3. Coating buffer: 0.05M, pH9.6 carbonate buffer;

[0062] 4. Washing solution: Each 1 liter washing solution is prepared as follows: 8.0g NaCl, 0.2g KH 2 PO 4 , 2.96gNa 2 HPO 4 ·12H 2 O. 1 mL of Tween-20 was dissolved in water an...

Embodiment 2

[0118] Example 2. Application of the kit

[0119] The EDTA-Cu chelate standard solution prepared above was diluted with the sample diluent to the following different Cu ion concentrations: 10ng / mL, 5ng / mL, 2.5ng / mL, 1.25ng / mL, 0.625ng / mL, 0.31ng / mL, 0.156ng / mL, 0.078ng / mL and 0.039ng / mL.

[0120] (1) Antibody coating: 1 mg / mL monoclonal antibody was diluted at 1:2000 and added to the ELISA plate, 100 μL per well, incubated at 37°C for 3 hours; poured out the solution in the ELISA plate, washed with Wash the plate 4 times with liquid and spin dry;

[0121] (2) Add the above EDTA-Cu chelate standard solution (experimental well) with different concentrations to the ELISA plate in step (1), 50 μL per well, and no EDTA-Cu chelate standard solution is added to the control well solution and add 50 μL of sample diluent;

[0122] (3) Add 0.5 μg / mL enzyme labeling substance to the above experimental wells and control wells, 50 μL per well; incubate at 37°C for 30 min; pour off the so...

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Abstract

The invention discloses an enzyme label and application thereof. The heavy metal enzyme label is a conjugate consisting of a complex and an enzyme, wherein the complex is a complex consisting of a conjugate of a chelant and a carrier protein and heavy metal. A method for preparing a copper-ion enzyme label has the advantages of simpleness, low synthesis cost and low HRP activity loss. The ELISA detection method constructed by the enzyme label has extremely high sensitivity. The method for preparing the enzyme label for copper ion ELISA detection has important reference value in successfully preparing an enzyme label for ELISA detection of other heavy metals, and the copper-ion rapid immunological detection method constructed by the method has wide application prospect.

Description

technical field [0001] The invention relates to a heavy metal enzyme marker and its application. Background technique [0002] Heavy metals are important pollutants in the environment and agricultural products. They can accumulate in the body of animals and humans through the food chain, especially in the liver, which will bring serious harm to human health. Copper is one of the trace elements necessary for the growth of life, but high concentrations of copper will have toxic effects on life. Aquatic organisms are particularly sensitive to the toxic effects of copper ions. [0003] At present, the analysis methods of heavy metal copper mainly include atomic absorption spectrometry, inductively coupled plasma emission light / mass spectrometry, X-ray fluorescence spectrometry, potentiometric dissolution analysis method and biosensor detection method developed in recent years. However, the detection of copper ions by these methods requires expensive instruments and equipment, ...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/577C07K16/44C12N5/20C12R1/91
Inventor 王保民赵洪伟南铁贵谭桂玉曹振高巍王敏孙硕李召虎
Owner CHINA AGRI UNIV
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