Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Chlorfenapyr hapten, chlorfenapyr artificial antigen and chlorfenapyr antibody as well as preparation method and application thereof

An artificial antigen, chlorfenapyr technology, applied in chemical instruments and methods, animal/human proteins, serum albumin, etc., can solve the problem of increased identification difficulty, reduced antibody affinity and specificity, and unfavorable hapten molecular characteristics. structure exposure and other problems, to achieve the effects of high purity and yield, enhanced immunogenicity, and easy control of reaction conditions

Pending Publication Date: 2020-11-17
BEIJING KWINBON BIOTECH +1
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Liu Shunyu et al. used to reduce the nitrile group on the pyrrole ring of chlorfenapyr to an amino group, and then couple it to the carrier protein. In this way, the distance between the modified site and the characteristic structure of the chlorfenapyr molecule is relatively close, which is not conducive to the exposure of the characteristic structure of the hapten molecule. , making it more difficult for the animal's immune system to recognize the characteristic structure of the hapten molecule, and reduce the affinity and specificity of the antibody

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Chlorfenapyr hapten, chlorfenapyr artificial antigen and chlorfenapyr antibody as well as preparation method and application thereof
  • Chlorfenapyr hapten, chlorfenapyr artificial antigen and chlorfenapyr antibody as well as preparation method and application thereof
  • Chlorfenapyr hapten, chlorfenapyr artificial antigen and chlorfenapyr antibody as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0033] Take 0.46g of alkynyl chlorfenapyr and add 70mL of 1mol / L dilute hydrochloric acid to dissolve, add 0.16g of potassium permanganate while stirring, heat after stirring well, react at 60°C for 2h, after the reaction is completed, add 6mol / L NaOH solution to adjust the pH to 6 , add ethyl acetate 80mL for extraction, separate the water phase, add 60mL of water to the organic phase and wash once, separate the water phase, add anhydrous sodium sulfate to dry and evaporate to dryness, purify with a silica gel column, and use dichloromethane and methanol at a volume ratio of 10 : 1 mixed solvent elution separation to obtain chlorfenapyr hapten 0.37g.

Embodiment 2

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

[0036] Take 18 mg of the chlorfenapyr hapten prepared in Example 1, add 1 mL of tetrahydrofuran to dissolve, add 14.2 mg of N-hydroxysuccinimide (NHS) and 17.4 mg of carbodiimide (EDC), and react at room temperature for 3 hours to obtain a hapten solution Liquid A: take 50 mg of human serum albumin (HSA), add 0.05mol / L PB buffer to dissolve, and obtain liquid B; add liquid A dropwise to liquid B, react at 4°C for 12 hours, and dialyze and purify with 0.02mol / L PBS For 3 days, the medium was changed 3 times a day to obtain the chlorfenapyr artificial antigen coupled with human serum albumin, which was aliquoted and stored at -20°C.

Embodiment 3

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

[0039] Take 9 mg of the chlorfenapyr hapten prepared in Example 1, add 1 mL of N,N-dimethylformamide (DMF) to dissolve, add 7.8 mg of NHS and 9.7 mg of EDC, and react at room temperature for 3 hours to obtain hapten solution A; Ovalbumin (OVA) 50mg, add 0.05mol / LPB buffer solution to dissolve to obtain solution B; add solution A to solution B dropwise, react at 4°C for 12h, dialyze and purify with 0.02mol / L PBS for 3 days, change the solution every day Three times, the chlorfenapyr artificial antigen coupled with ovalbumin was obtained, aliquoted, and stored at -20°C.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Sensitivityaaaaaaaaaa
Sensitivityaaaaaaaaaa
Login to View More

Abstract

The invention discloses a chlorfenapyr hapten, an artificial antigen and an antibody as well as a preparation method and application thereof; the chlorfenapyr hapten provided by the invention not onlyretains the characteristic structure of chlorfenapyr to the greatest extent so that the immunogenicity of the chlorfenapyr hapten is obviously enhanced, but also has carboxyl capable of being coupledwith carrier protein; therefore, the chlorfenapyr artificial antigen obtained by coupling the chlorfenapyr hapten and the carrier protein is used for immunizing animals, animal immune response can bestimulated to generate an antibody with higher specificity and sensitivity, the sensitivity of the chlorfenapyr antibody can reach 1 microgram / L through detection, the cross reaction rate with otherpyrrole pesticides is low, and the sensitivity of the chlorfenapyr antibody is high. A basis is provided for subsequent establishment of various immunoassay methods of chlorfenapyr.

Description

technical field [0001] The invention belongs to the field of food safety detection. More specifically, the present invention relates to chlorfenapyr haptens, artificial antigens and antibodies and their preparation methods and applications. Background technique [0002] Chlorfenapyr (chlorfenapyr), also known as chlorfenapyr, is an arylpyrrole insecticide and acaricide with broad insecticidal spectrum, high efficacy, long duration, low dosage, no cross-resistance, It has the advantages of being safe to crops, and has both stomach poisoning and contact killing effects. It has been registered for use in various crops such as vegetables, fruit trees, and cotton. With the increase in the range and amount of chlorfenapyr used, its half-life in food and the environment is long, and its residual toxic and side effects are gradually emerging. Medical research has shown that chlorfenapyr can affect the DNA of liver, kidney, spleen cells of mice and the reproductive performance of w...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07D207/34C07K14/765C07K14/77C07K14/795C07K16/44G01N33/53G01N33/577
CPCC07D207/34C07K14/765C07K14/77C07K14/795C07K16/44G01N33/577G01N33/5308G01N2430/10Y02P20/55
Inventor 何方洋万宇平崔廷婷刘玉梅崔娜凡静静张坤
Owner BEIJING KWINBON BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com