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Method for detecting existence of heavy metal copper ion in sample and special kit thereof

A technology for the detection of samples and metal ions, which is applied in the directions of microorganism-based methods, biological tests, material inspection products, etc., can solve the problems of inability to use rapid detection, high detection cost, time-consuming, etc., and achieves improved ELISA detection sensitivity and synthesis cost. The effect of low and broad application prospects

Inactive Publication Date: 2011-02-09
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Method for detecting existence of heavy metal copper ion in sample and special kit thereof
  • Method for detecting existence of heavy metal copper ion in sample and special kit thereof
  • Method for detecting existence of heavy metal copper ion in sample and special kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045]Embodiment 1, composition and preparation of kit

[0046] 1. The composition of the kit

[0047] 1. Coating agent: a complex formed by a conjugate of p-aminobenzylethylenediaminetetraacetic acid and BSA and cobalt ions (cobalt-p-aminobenzyl-EDTA-BSA).

[0048] 2. Anti-copper-EDTA chelate monoclonal antibody: the monoclonal antibody secreted by the hybridoma cell line Cu-EDTA 6A9 with the deposit number CGMCC No.3987;

[0049] 3. Coating buffer: 0.05M, pH9.6 carbonate buffer;

[0050] 4. Washing solution: Prepare each 1 liter of washing solution as follows: mix 8.0g NaCl, 0.2g KH 2 PO 4 , 2.96gNa 2 HPO 4 12H 2 O. Dissolve 1 mL of Tween-20 in water and make up to 1 L with water.

[0051] 5. Sample diluent: Prepare the sample diluent as follows: Dissolve 1 mL of Tween 20 and 1 g of gelatin in 1 L of PBS buffer to obtain the sample diluent;

[0052] Each liter of PBS buffer is composed of NaCl, KH 2 PO 4 、Na 2 HPO 4 12H 2 Composed of O and water; the concentrati...

Embodiment 2

[0107] Embodiment 2, the sensitivity test of kit

[0108] Dilute the EDTA-Cu chelate standard solution prepared above with the sample diluent to the following different concentrations: 20ng / mL, 10ng / mL, 5ng / mL, 2.5ng / mL, 1.25ng / mL, 0.625ng / mL mL, 0.31 ng / mL, 0.156 ng / mL, and 0.078 ng / mL.

[0109] (1) Coating of the original coating: the cobalt-p-aminobenzyl EDTA-BSA antigen prepared above was diluted 1:4000 and added to the microtiter plate, 100 μL per well, incubated at 37°C for 3 hours; For the solution in the microplate plate, wash the plate 4 times with washing solution, and spin dry;

[0110] (2) Add the above-mentioned different concentrations of EDTA-Cu chelate standard solution (experimental well) to the microtiter plate of step (1), 50 μL per well, and do not add EDTA-Cu chelate standard in the control well Solution and add 50μL sample diluent;

[0111] (3) Add 0.25 μg / mL monoclonal antibody to the above-mentioned experimental wells and control wells, 50 μL per wel...

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PUM

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Abstract

The invention discloses a method for detecting the existence of a heavy metal copper ion in a sample and a special kit thereof. The kit comprises a coating antigen and an antibody resistant to heavy metal to be detected, wherein the coating antigen and the antibody resistant to the heavy metal to be detected interact with each other; and the affinity between the coating antigen and the antibody resistant to the heavy metal to be detected is less than the affinity between the heavy metal to be detected and the antibody resistant to the heavy metal to be detected. Experiments prove that the preparation method of the coating antigen is simple and has clear synthesis steps, low synthesis cost and good effect in an unequal competition enzyme-linked immuno sorbent assay (ELISA) detection method for preparing copper ions provided by the invention. The ELISA detection sensitivity of the coating antigen prepared by the method of the invention is enhanced remarkably. The method for preparing the coating antigen and a copper ion rapid immunological detection method established by the method have wide application prospects.

Description

technical field [0001] The invention relates to a method for detecting whether a sample contains heavy metal copper ions and a special kit thereof. 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. Copper is one of the trace elements necessary for the growth of living organisms, but high concentrations of copper will have a toxic effect on living organisms. [0003] At present, the analysis methods of heavy metal copper 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 copper ions by these methods requires expensive instruments and equipment, high detection costs, time-consuming, and cannot be used for on-site rapid dete...

Claims

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Application Information

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IPC IPC(8): C12R1/91G01N33/566G01N33/53C07K16/44C12N5/20
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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