Clothianidin antigen, antibody and application thereof

A clothianidin and antibody technology is applied in the field of immunochemical analysis to achieve the effects of large analysis capacity, high accuracy, and easy popularization and promotion.

A clothianidin and antibody technology is applied in the field of immunochemical analysis to achieve the effects of large analysis capacity, high accuracy, and easy popularization and promotion.

CN102875671AInactive Publication Date: 2013-01-16NANJING AGRICULTURAL UNIVERSITY
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  • Clothianidin antigen, antibody and application thereof
  • Clothianidin antigen, antibody and application thereof
  • Clothianidin antigen, antibody and application thereof

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Embodiment Construction

[0025] 1 Synthesis of artificial hapten

[0026] Substituting the chlorine atom on the thiazole ring of the clothianidin molecule to synthesize a hapten with a four-atom connecting arm, which not only retains the characteristic group of clothianidin to the greatest extent, but also forms a carboxyl terminal group, which can better follow the protein coupling. The product was purified by mass spectrometry (ESI) and H NMR spectroscopy ( 1 H-NMR) identification. The chemical name of clothianidin artificial hapten is 3-(5-((3-methyl-2-nitroguanidine)-ylmethyl)thiazole-2-mercapto)propionic acid, and its molecular structure is as follows:

[0027]

[0028] 1.1 Synthesis of artificial haptens

[0029] Dissolve 8mmol (0.45g) of KOH in 20mL of ethanol, add 4mmol (0.42g) of β-mercaptopropionic acid and stir, after complete dissolution, add 4mmol (1.02g) clothianidin, stir and reflux at 80°C for 2h. After the reaction, the reaction solution was filtered, and the filtrate was conce...

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Abstract

The invention relates to a clothianidin antigen, an antibody and an application thereof, and belongs to the technical field of immunochemistry analysis. The clothianidin antigen and the antibody of the invention are specially used for clothianidin specific polyclonal antibody preparation, enzyme-linked immunoassay (ELISA), and high-sensitivity and rapid detection of clothianidin residues in environment and agricultural products. A hapten is synthesized by substituting a chlorine atom on a thiazole ring of clothianidin, has a chemical name of 3-(5-((3-methyl-2-nitroguanidine)-yl methyl)thiazole-2-mercapto)propionic acid, and is coupled with bovine serum albumin and ovalbumin to prepare an antigen and an envelope antigen. Newzealand white rabbit is immunized by the immunizing antigen to obtain the specific polyclonal antibody of clothianidin. The established ELISA linear scope is 1.1 microgram / L-2 mg / L, and the detection limit is 1.1 microgram / L. The hapten synthetic technology of the invention is simple and feasible, high in antibody specificity, and suitable for detection and on-site monitoring of mass samples in environment and agricultural products.

Description

1. Technical field [0001] The invention relates to clothianidin antigen, antibody and application thereof, and belongs to the technical field of immunochemical analysis. It is specially used for the preparation of clothianidin polyclonal antibody, and the enzyme-linked immunosorbent assay method (ELISA) for clothianidin residues in the environment and agricultural products. 2. Background technology [0002] Clothianidin, chemical name 1-(2-chloro-1,3-thiazol-5-ylmethyl)-3-methyl-2-nitroguanidine, jointly developed by Sumika Takeda / Bayer, It was registered in Japan in December 2001. The chemical structure is shown in the figure below. Molecular formula: C 6 h 8 CIN 5 o 2 S, relative molecular mass: 249.68, melting point: 176.8°C. Clothianidin is a new type of nicotinic insecticide containing a thiazole ring, which mainly acts on the nicotinic acetylcholine receptors in the post-synaptic membrane of the insect nervous system. Clothianidin is widely used in the prevention...

Claims

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Application Information

Patent Timeline
16 Jan 2013
Publication
CN102875671A
IPC
C07K16/06; G01N33/53
Inventors
王鸣华; 李明