Preparation of thiacloprid monoclonal antibodies and ELISA detection method

A monoclonal antibody and detection method technology, applied in the field of immunology, can solve the problems that the thiacloprid immunoassay method has not been reported yet, and achieve high affinity and strong specificity

Inactive Publication Date: 2015-10-14
ZHEJIANG MARINE DEV RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there have been many enzyme-linked immunoassay reports for nicotinic

Method used

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  • Preparation of thiacloprid monoclonal antibodies and ELISA detection method
  • Preparation of thiacloprid monoclonal antibodies and ELISA detection method
  • Preparation of thiacloprid monoclonal antibodies and ELISA detection method

Examples

Experimental program
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Embodiment Construction

[0023] Below in conjunction with the examples, the specific implementation of the present invention will be further described in detail. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0024] 1. Synthesis of haptens

[0025] Dissolve 1.12g (4mmol) of thiacloprid, 0.42g (4mmol) of β-mercaptopropionic acid and 0.45g (8mmol) of KOH in 20ml of DMSO, and at the same time slowly raise the temperature to 100°C, keep stirring at this temperature for 2 hours; Finally, 50 mL of water was added to cool the reaction mixture to room temperature, adjusted to pH=3.0 with 2M hydrochloric acid and extracted with dichloromethane (3×30 mL), the organic phase was washed with water (3×30 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure, After silica gel column chromatography, a white powder was obtained.

[0026] 2. Preparation of artificial antigen

[0027] Preparation of im...

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Abstract

The invention discloses preparation of thiacloprid monoclonal antibodies. The preparation method comprises the steps of synthesizing thiacloprid haptens, coupling the thiacloprid haptens with bovine serum albumin to prepare immunizing antigens, immunizing a BALB/c mouse, fusing the spleen cells of the immunized mouse with Sp2/0 myeloma cells, and performing screening and cloning to obtain hybrid strains of continuous and stable secreting thiacloprid monoclonal antibodies. The thiacloprid monoclonal antibodies have high affinity to thiacloprid and strong specificity, and do not have cross reaction with nicotine insecticide similar to acetamiprid, imidacloprid and the like.

Description

Technical field: [0001] The invention belongs to the technical field of immunology, and more specifically relates to the preparation and detection method of thiacloprid monoclonal antibody. Background technique: [0002] Thiacloprid is a new type of chlorinated neonicotinoid insecticide, jointly developed by Bayer Agrochemical Company of Germany and Bayer Agrochemical Company of Japan, which has special effects on piercing-sucking and chewing mouthparts pests. The mechanism of action is different from other traditional insecticides. It mainly acts on the post-junction membrane of insect nerves. By binding to nicotinic acetylcholine receptors, it interferes with the normal conduction of the insect nervous system, causing blockage of nerve channels, resulting in a large accumulation of acetylcholine. As a result, the insect is extremely excited, and the whole body convulses, paralyzes and dies. With strong contact, stomach poisoning and systemic effects, thiacloprid has no cr...

Claims

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Application Information

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IPC IPC(8): C07K16/44C07K14/765C07D417/06G01N33/577
Inventor 相兴伟
Owner ZHEJIANG MARINE DEV RES INST
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