Monoclonal antibody for detecting imidacloprid pesticide residue

A monoclonal antibody, imidacloprid technology, applied in the biological field, can solve the problems such as late, only in recent years, relevant researchers have begun to pay attention, and achieve the effect of eliminating the pretreatment process, good specificity, and large analysis capacity.

Inactive Publication Date: 2010-11-10
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Foreign researchers have successively reported the immunological residue detection methods of various pesticides in the 1990s. Domestic research in this area is relatively late. Only in recent yea

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: Detection of imidacloprid pesticide residues in tap water samples

[0057] Main conditions of ELISA reaction system

[0058] Add 50 μL of H2-OVA coating material 1.82 μg / mL to each well of the microanalysis plate, dilute the monoclonal antibody 32 times as the working concentration, use 1% gelatin as the blocker, and antigen-antibody reaction solution (phosphoric acid containing 0.05% Tween 20) The content of acetone in salt buffer solution) is 5%, the pH value is 7.5, the ionic strength is 0.8 mol / L, and 50 μL is placed in each well of the microanalysis plate.

[0059] Sample preparation for testing

[0060] Tap water sample: Add 250 mg net weight of imidacloprid standard substance into 1 mL of acetone solution to dissolve, and prepare 250 mg / mL imidacloprid acetone solution, measure 1 L of tap water, add 1 μL 250 mg / mL imidacloprid acetone solution, and prepare 250 μg / L imidacloprid tap water sample.

[0061] The above samples were detected by ELISA dete...

Embodiment 2

[0085] Example 2: Detection of imidacloprid pesticide residues in rice samples

[0086] The main conditions of the ELISA reaction system are the same as in Example 1.

[0087] Sample preparation for testing

[0088] Weigh 1g of rice, grind it into powder, add 1mL of imidacloprid standard solution (dissolved in acetone) with a concentration of 250μg / L to 1g of the sample, then the content of imidacloprid in the rice sample is 250μg / kg, shake (60r / min) 2 After shaking for several hours, let it stand for a few minutes, take 100 μL of the supernatant and add it to 900 μL of phosphate buffer containing 0.05% Tween 20 to prepare the solution to be tested.

[0089] The above samples were detected by ELISA detection method, and the operation was as follows:

[0090] Steps (1), (2) are the same as in Example 1.

[0091] (3) Add the mixture of antibody and sample to be tested

[0092] The 16-fold diluted cell culture supernatant was fully mixed with the test solution and phosphate b...

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PUM

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Abstract

The invention relates to an imidacloprid pesticide against monoclonal antibody and a preparation method thereof, and belongs to the technical field of biology. The preparation method comprises the following steps of: immunizing a BALB/c mouse by using a coupling substance of immune hapten 1-[6-(2-carboxyethylsulfenyl-3-pyridine) methyl]-N-nitro-2-imidazoline imine and bovine serum albumin, preparing hybrid tumor cells from spleen cells and myeloma cells Sp2/0 of the immunized mouse by the hybrid tumor technology, and obtaining hybrid tumor strains 2F11/A9 capable of stably secreting the imidacloprid pesticide against monoclonal antibody. By effectiveness verification, the antibody can be used for sensitive and quick detection of imidacloprid residues in agricultural production environments and agricultural products. The preparation technique for the imidacloprid pesticide against monoclonal antibody is simple and feasible, does not need special instruments and equipment in the whole preparation process of the antibody, and is easy for scale production in factories.

Description

(1) Technical field [0001] The invention relates to an anti-pesticide imidacloprid monoclonal antibody and a preparation method thereof, belonging to the field of biotechnology. Dedicated to the sensitive and rapid detection of imidacloprid residues in agricultural products and agricultural production environments, especially suitable for the detection of large batches of samples. (2) Background technology [0002] Imidacloprid, 1-(6-chloro-3-pyridylmethyl)-N-nitro-2-imidazolidinyl imine, belongs to a new type of neonicotinoid insecticide, moderate toxicity, through the action of acetylcholine in insects esterase receptors to paralyze the pest and eventually die. Imidacloprid can effectively control agricultural pests such as aphids, planthoppers, and leafhoppers, and has been widely used in agricultural production. Due to the extensive use of this pesticide in agricultural production in recent years, it not only pollutes the environment, but also poses a serious threat to...

Claims

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Application Information

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IPC IPC(8): C07K16/44C07K14/765C12N5/20G01N33/577C12R1/91
Inventor 刘凤权李刚钱国良华修德秦娜
Owner NANJING AGRICULTURAL UNIVERSITY
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