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Chlordane hapten, artificial antigen and antibody, preparation method and application thereof

A technology of artificial antigens and haptens, applied in chemical instruments and methods, preparation of cyanide reaction, preparation of organic compounds, etc.

Active Publication Date: 2020-10-27
BEIJING KWINBON BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the chlordane hapten in China

Method used

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  • Chlordane hapten, artificial antigen and antibody, preparation method and application thereof
  • Chlordane hapten, artificial antigen and antibody, preparation method and application thereof
  • Chlordane hapten, artificial antigen and antibody, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A preparation method of chlordane hapten, the steps are as follows:

[0033] Take 0.423g of chlordane oxide and add 100mL of methanol to dissolve; take 0.206g of aminobutyric acid and add 5mL of water to dissolve, add it to the methanol solution of chlordane oxide, and stir thoroughly; take 0.43g of potassium carbonate and add 5mL of water to dissolve, add it to the reaction solution, Stir at ℃ for 5 hours, stop the reaction, add 6mol / L hydrochloric acid to adjust the pH to 6, concentrate by rotary evaporation to remove methanol, add 50mL of water, 100mL of chloroform, extract three times, combine the organic phases, evaporate to dryness to obtain a yellow oil, add 12mL of absolute ethanol to reweight Crystallization gives the chlordane hapten.

Embodiment 2

[0035] A preparation method of chlordane artificial antigen, the steps are as follows:

[0036] Take 19.6 mg of the chlordane hapten prepared in Example 1, add N, N-dimethylformamide (DMF) 1 mL to dissolve, add N-hydroxysuccinimide (NHS) 13.3 mg, carbodiimide (EDC) 15.7 mg, react at room temperature for 3 hours to obtain hapten solution A; take bovine serum albumin (BSA) 50 mg, add 0.05mol / L CB buffer to dissolve, and obtain B liquid; add A dropwise to B liquid, 4 ℃ After reacting for 12 hours, dialyze and purify with 0.02mol / L PBS for 3 days, and change the medium 3 times a day to obtain artificial chlordane antigen coupled with bovine serum albumin, which is aliquoted and stored at -20°C.

Embodiment 3

[0038] A preparation method of chlordane artificial antigen, the steps are as follows:

[0039] Take 12 mg of the chlordane hapten prepared in Example 1, add 1 mL of dimethyl sulfoxide (DMSO) to dissolve, add 8.4 mg of NHS, 10.2 mg of EDC, and react at room temperature for 3 hours to obtain hapten solution A liquid; take ovalbumin (OVA ) 50mg, add 0.05mol / L CB buffer solution to dissolve to obtain liquid B; add liquid A dropwise to liquid B, react at 4°C for 12h, dialyze and purify with 0.02mol / L PBS for 3 days, change the liquid 3 times a day, and obtain Chlordane artificial antigen coupled with ovalbumin, aliquoted and stored at -20°C.

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Abstract

The invention discloses a chlordane hapten, an artificial antigen and an antibody, a preparation method and application thereof. The chlordane hapten provided by the invention retains the characteristic structure of chlordane to the greatest extent, so that the immunogenicity of the chlordane hapten is obviously enhanced, and the chlordane hapten has carboxyl capable of being coupled with carrierprotein; a chlordane hapten is coupled with a carrier protein to obtain a chlordane artificial antigen for immunizing animals, an antibody with stronger specificity and higher sensitivity is more favorably generated by stimulating animal immune response, the sensitivity of the chlordane antibody can reach 0.1microg / L through detection, the cross reaction rate of the chlordane antibody and other cyclopentadiene organochlorine pesticides is low, and a basis is provided for subsequent establishment of various immune analysis methods of chlordane.

Description

technical field [0001] The invention belongs to the field of food safety detection. More specifically, the present invention relates to chlordane haptens, artificial antigens and antibodies and their preparation methods and applications. Background technique [0002] Chlordane is a broad-spectrum organochlorine insecticide, aliased as octachloro-hexachloro-methyleneindene, which is often used to kill underground pests, such as mole crickets, cutworms, rice grass pests, etc., and has a significant effect on termite control . Chlordane has contact and stomach toxicity to insects, has no phytotoxicity to plants at insecticidal concentrations, and is toxic to mammals. Chlordane toxicity is mainly manifested in causing biological endocrine system disorders, destroying reproductive and immune systems, and inducing cancer and neurological diseases. It is a serious hazard to the environment. It can penetrate into the groundwater system through the surface soil and cause pollution...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C229/14C07C227/18C07C227/40C07K14/765C07K14/77C07K14/795C07K16/44G01N33/53G01N33/68
CPCC07C229/14C07C227/18C07C227/40C07K14/765C07K14/77C07K14/795C07K16/44G01N33/5308G01N33/6854C07C2603/66C07K2317/33C07K2317/92G01N2430/10Y02P20/55
Inventor 崔海峰何方洋崔廷婷王兆芹屈秀玲崔娜宋灏万宇平
Owner BEIJING KWINBON BIOTECH
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