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Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl

A technology of methyl parathion and monoclonal antibody, applied in the biological field, can solve the problems such as unreported immunological multi-residue detection method, and achieve the effects of easy popularization and promotion, high cost and wide detection range.

Active Publication Date: 2014-07-30
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, gas chromatography (GC) and high performance liquid chromatography (HPLC) are used for the detection of these two pesticide residues in agricultural products; The immunological multi-residue detection method for pesticides has not been reported yet

Method used

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  • Bispecific monoclonai antibody capable of simultaneously detecting imidacloprid and parathion-methyl

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Imidacloprid, methyl parathion residue detection in tap water samples

[0038] Main conditions of ELISA reaction system

[0039] The original coating concentration of imidacloprid was 4.88ug / ml, and the dilution ratio of ascites was 2000 times; the original coating concentration of methyl parathion was 2.3ug / ml, and the dilution ratio of ascites was 4000 times. 1% gelatin is used as the blocking solution, the methanol content in the antigen-antibody reaction solution (phosphate buffer containing 0.05% Tween 20) is 5%, the pH value is 8.5, and the ionic strength is 0.8mol / L. 50ul.

[0040] Sample preparation for testing

[0041] Tap water sample: imidacloprid and methyl parathion pesticides were formulated into methanol solutions of different concentrations, the concentrations of imidacloprid were 6, 12ug / ml, and the concentrations of methyl parathion were 0.64, 1.28ug / ml. After the above pesticide methanol solution was diluted 20 times with tap water, the p...

Embodiment 2

[0053] Imidacloprid, methyl parathion residue detection in embodiment 2 pond water sample

[0054] ELISA reaction system main condition is the same as embodiment 1

[0055] Sample preparation for testing

[0056] Pond water samples: Centrifuge the collected pond water and take the supernatant for the experiment. Imidacloprid and methyl parathion pesticides were formulated into methanol solutions with different concentrations, the concentrations of imidacloprid were 6, 12ug / ml, and the concentrations of methyl parathion were 0.64, 1.28ug / ml. After diluting the above pesticide methanol solution 20 times with pond water, the pesticide concentrations are respectively: imidacloprid 300, 600ng / ml, methyl parathion 32, 64ng / ml.

[0057] The above samples were detected by ELISA detection method, and the operation was as follows:

[0058] Step (1), (2) are the same as embodiment 1

[0059] (3) Sample addition

[0060] Mix the above-prepared imidacloprid, methyl parathion solution ...

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Abstract

The invention relates to a bispecific monoclonai antibody capable of simultaneously recognizing imidacloprid and parathion-methyl pesticides, and a preparation method thereof, belonging to the field of biological technology. The bispecific monoclonai antibody FQ-D6 which can be stably secreted and simultaneously resist imidacloprid and parathion-methyl pesticides is prepared by fusing hybridoma cell capable of secreting anti-parathion-methyl monoclonal antibody and an imidacloprid antigen-immunized rat splenocyte cell through a hydridizatio-hybridoma technique. Proved by effectiveness verification, the antibody can be used for fast and sensitively detecting residuals of the parathion-methyl and the imidacloprid in agricultural production environment and agricultural products. The preparation technology of the bispecific monoclonai antibody capable of simultaneously resisting imidacloprid and parathion-methyl pesticides is simple, convenient and feasible, the whole preparation process of the antibody is not required for special instruments and equipment, thereby being easy for factorization scale production.

Description

(1) Technical field [0001] The invention relates to a bispecific monoclonal antibody applied to simultaneous detection of imidacloprid and methyl parathion and a preparation method thereof, belonging to the field of biotechnology. Dedicated to the sensitive and rapid detection of imidacloprid and methyl parathion residues in agricultural products and agricultural production environments, especially for the detection of large batches of samples. (2) Background technology [0002] Imidacloprid (1-(6-chloro-3-pyridylmethyl)-N-nitro-2-imidazolidinium) and parathion-methyl (Parathion-methyl, O, O-dimethyl-O -(4-nitrophenyl)phosphorothioate) is an insecticide widely used in agricultural production, imidacloprid is a new type of neonicotinoid insecticide with moderate toxicity, and methyl parathion is a highly toxic organophosphate class of pesticides. At present, gas chromatography (GC) and high performance liquid chromatography (HPLC) are used for the detection of these two pes...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/44C12N5/20C12R1/91
Inventor 刘凤权王利民华修德李刚
Owner NANJING AGRICULTURAL UNIVERSITY
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