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Preparing method of heavy metal cadmium monoclonal antibody

A monoclonal antibody, metal chelator technology, applied in the direction of anti-animal/human immunoglobulin, measuring device, instrument, etc., can solve the problems of late start, no research report, etc., achieve good stability and easy factoryization The effect of large-scale production and strong practicability of antigens

Inactive Publication Date: 2007-08-15
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Our country started relatively late in this area, and only in recent years have units and researchers paid attention to and engaged in the research of heavy metal immune rapid detection technology. At present, there is no research report on this kind of method in China.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The detection of embodiment 1 cadmium standard solution (Sigma Co.)

[0037] The indirect competitive ELISA method was used to conduct a preliminary exploration of the detection curve. Methods as below:

[0038] Coating: Dilute the coating antigen Cd-1-(4-isothiocyanatophenyl)-ethylenediaminetetra Acetic acid-bovine serum albumin, 50 μl / well was added to a 96-well enzyme-linked micro-assay plate (Coring Co.), and left overnight at 4°C.

[0039] Washing: 5 times with PBST

[0040] Blocking: PBST diluted gelatin to 1%, 100 μl / well, 37°C for 1.5 hours.

[0041] Washing: 5 times with PBST

[0042] Adding samples: Dilute the cadmium standard solution 10 times step by step with phosphate buffer containing 2mM ethylenediaminetetraacetic acid, place the diluted solution at room temperature for 30 minutes, then mix it with an equal volume of cadmium antibody diluted to a certain multiple and react at room temperature for 45 minutes. Add 50 μl / well to the enzyme-linked plate...

Embodiment 2

[0049] Detect cadmium residue in the tap water sample of embodiment 2

[0050] Cd-1-(4-isothiocyanatophenyl)-ethylenediaminetetraacetic acid-bovine serum albumin 0.25 μg / ml coated microanalysis plate 50 μl / well, cadmium antibody supernatant diluted 50 times as the working concentration, 1% gelatin was used as a blocker, the pH value of the antigen-antibody reaction matrix (HBS) was 7.0, and the ionic strength was 0.15 mol / L, 50 μl / well in the microanalysis plate.

[0051] Sample preparation to be tested:

[0052] Tap water sample: measure 100 μl of tap water, add 0.5 μl of 1000ug / ml cadmium standard solution, and prepare a 5ug / ml liquid sample.

[0053] Using cadmium ELISA detection method to detect the above samples, the operation is as follows:

[0054] 1) Coating:

[0055] Add 50 μl of Cd-1-(4-isothiocyanatophenyl)-ethylenediaminetetraacetic acid-bovine serum albumin 0.25 μg / ml to each well of a microanalysis plate, and place in a refrigerator at 4°C overnight or for 2 h...

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Abstract

The invention discloses a making method of heavy-metal cadmium monoclonal antibody in the biological technical domain, which is characterized by the following: detecting residual cadmium in the agricultural product sensitively and rapidly; coupling cadmium ion and carrier protein through 1-(4-isothiocyanic phenyl)-ethanediamine tetraacetate difunctional metal chelant to form complete antigen; obtaining stable Cd-EDTA monoclonal antibody.

Description

(1) Technical field [0001] The invention relates to the preparation of heavy metal cadmium monoclonal antibody and belongs to the field of biotechnology. It is specially used for the preparation of monoclonal antibodies that specifically recognize the heavy metal cadmium, and the highly sensitive and rapid detection of heavy metal residues in agricultural products and agricultural production environments, especially suitable for large-scale sample detection and on-site monitoring. (2) Background technology [0002] The main pollution sources of cadmium in the environment are: waste water and waste gas produced during the mining, beneficiation and smelting of lead-zinc mines; the production and processing of alloy steel; the production waste water of cadmium electroplating, dyes, pesticides, paints, glass, ceramics, Production and processing of photographic materials, etc. Cadmium residues in the environment are gradually enriched through the food chain, and a very small amo...

Claims

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Application Information

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IPC IPC(8): C07K16/18G01N33/531
Inventor 刘凤权朱晓霞
Owner NANJING AGRICULTURAL UNIVERSITY
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