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Immunological method for quickly detecting heavy metal lead ions and kit

A lead ion and immunology technology, which is applied in the field of immunology and kits for rapid detection of heavy metal lead ions, can solve the problems of difficult product production and high cost, and achieve the effect of reducing manufacturing cost and retaining sensitivity and accuracy

Active Publication Date: 2013-08-21
JINAN UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

IC-ELISA is used to detect heavy metal lead, although it can achieve a high detection level, but its cost is too high (requires excessive expensive chelating agent), and it is difficult to be really used in product production

Method used

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  • Immunological method for quickly detecting heavy metal lead ions and kit
  • Immunological method for quickly detecting heavy metal lead ions and kit
  • Immunological method for quickly detecting heavy metal lead ions and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Detect heavy metal lead ion with immunological method of the present invention:

[0030] (1) Preparation of monoclonal antibody solution:

[0031] ①Synthesis of artificial antigen

[0032] Keyhole limpet hemocyanin (KLH), which is far from mouse, was selected as the immune carrier, and ovalbumin (OVA) was selected as the detection carrier.

[0033] 1) Mother liquor equipment:

[0034] Solution A: Weigh 12mg of CHX-A”-DTPA (diethyltriaminepentaacetic acid) and dissolve it in 1ml of pure water, heat to 60°C, and dissolve;

[0035] Solution B: Weigh 17mg Pb(NO 3 ) 2 Dissolve in 200 μl of 2% dilute nitric acid by volume;

[0036] Solution C: Weigh 10mg OVA, dissolve in 2ml HEPES (10mM, pH9.0), and place at 4°C;

[0037] Solution D: Measure 850μl KLH (5.8mg·ml -1 ), and stored at 4°C for later use.

[0038] 2) Preparation of immune antigen:

[0039] Take 30 μl of step 1) B solution and add dropwise to 350 μl of step 1) A solution, shake and react at room temperature...

Embodiment 2

[0064] Detect the specificity of the immunological method of the present invention:

[0065] The lead ion standard solution among the embodiment 1 is changed into Pb of different concentrations, Cd, Hg, Cr, Ca, Cu, Fe, heavy metal standard solutions such as Mn (purchased from National Iron and Steel Material Testing Center Iron and Steel Research Institute), adopt the same method operation, and read the final OD 450 value to test the specificity of the system for detecting lead. Test results such as figure 2 As shown, this result shows that the antibody is specific for lead ions, and there is no cross-reaction with other heavy metal ions.

Embodiment 3

[0067] Draw the lead ion ELISA detection system standard curve:

[0068] Add different concentrations of lead ion standard solutions (μg / L) on the antigen CHX-A"-DTPA-OVA coated plate with a concentration of 100ng / well: 10 -4 、10 -3 、10 -2 、10 -1 、10 0 、10 1 、10 2 、10 3 . After the lead ion solution reacted with the antigen at 37°C, add 100 ng / well of anti-lead monoclonal antibody at a certain concentration; after the reaction is completed, add the enzyme-labeled secondary antibody IgG-HRP; finally add TMB substrate for color development (the specific operation is the same as in the examples) 1). Sent to a microplate reader to read the absorbance value at 450nm (OD450). Get the logarithm with lead standard solution concentration as abscissa, OD450 value as ordinate, select the linear segment in the test result, draw and detect lead standard curve (such as image 3 shown). It can be concluded from the results that the system can effectively detect lead ions in the ra...

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Abstract

The invention discloses an immunological method for quickly detecting heavy metal lead ions and a kit. The immunological method is an indirect enzyme-linked immunological technology, and comprises the following steps of: coating lead ions in a lead-free antigen combined sample through artificially synthesized antigens to form lead-containing antigens; reacting the lead-containing antigens and anti-lead monoclonal antibodies to form an antigen-lead-antibody structure chain; reacting the structure chain and enzyme labeled secondary antibodies to form an antigen-lead-antibody-secondary antibody structure chain; and performing color development reaction, and judging the lead ion content of the detected sample according to the color development degree. The kit for implementing the immunological method comprises an artificial antigen coated and closed enzyme-linked immunosorbent assay (ELISA) plate and the anti-lead monoclonal antibodies, wherein the artificial antigen means CHX-A'-DTPA-OVA; and the anti-lead monoclonal antibodies are generated by monoclonal cell strains with collection date of August 25th, 2010, collection number of CCTCC C201084 and name of 6H8 / 4F7. The indirect enzyme-linked immunological detection technology is applied in small molecular detection for the first time, and the detection effect is good.

Description

technical field [0001] The invention particularly relates to an immunological method and kit for rapid detection of heavy metal lead ions, which can be used for rapid detection of heavy metal lead ion residues in environmental agricultural products. Background technique [0002] In recent years, due to the massive release of industrial wastewater and mine development waste, the residues of heavy metal pollutants in soil and water in my country have increased rapidly, and have directly affected the quality and safety of agricultural and aquatic products. Heavy metal elements can be enriched in animals and plants and finally transferred to the human body through the food chain. Human poisoning incidents occur frequently due to accidental consumption of agricultural and aquatic products containing residues of heavy metal pollutants. This also shows that we urgently need long-term and effective monitoring of the quality of environmental agricultural products and aquatic product...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53G01N33/543G01N33/577G01N21/78
Inventor 江天久包财秦超王晔
Owner JINAN UNIVERSITY
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