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

Phenylethanolamine A hapten, antigen as well as preparation method and application thereof

A phenylethanolamine and hapten technology, which is applied in the fields of phenethanolamine A hapten, monoclonal antibody preparation, and antigen, can solve the problems of complicated pretreatment, unsuitable for screening of a large number of samples and operation by grass-roots personnel, and time-consuming, and achieves the preparation method. Simple and feasible, high yield, low cost effect

Inactive Publication Date: 2016-01-27
北京维德维康生物技术有限公司
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the method for detecting phenylethanolamine A is mainly an instrument method——high performance liquid chromatography mass spectrometry. This method requires professional equipment and professional personnel to operate, and the pretreatment is complicated and time-consuming. It is not suitable for the screening and screening of a large number of samples. Grassroots personnel operation

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
  • Phenylethanolamine A hapten, antigen as well as preparation method and application thereof
  • Phenylethanolamine A hapten, antigen as well as preparation method and application thereof
  • Phenylethanolamine A hapten, antigen as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1, preparation phenylethanolamine A hapten

[0031] Phenylethanolamine A138mg is dissolved in 10ml1mol / LHCl and 4ml dimethyl sulfoxide; add

[0032] Zinc powder 100mg, stirred and reacted in 80°C water bath for 30min, cooled and filtered, the filtrate was adjusted to pH 8.0, extracted twice with 20ml ethyl acetate, combined the organic phase, and dried in vacuo; the residue was dissolved in 10ml DMF, and 60mg potassium carbonate and 82mg iodoacetic acid were added, Heat the reaction at 90°C for 3 hours, lower it to room temperature, add 50ml of distilled water, adjust the pH to 6.0 with 1M hydrochloric acid, extract twice with 50ml of ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, filter, and vacuum-dry the filtrate, the residue is half antigen.

[0033] The structural formula of the phenylethanolamine A hapten is shown in formula I.

Embodiment 2

[0034] Embodiment 2, preparation and identification of phenylethanolamine A artificial antigen

[0035] 1. Synthesis of phenylethanolamine A immune antigen

[0036] Weigh 10mg of hapten and dissolve in 2ml DMF, add EDC4.3mg, NHS5.1mg, stir at room temperature

[0037] 50mg BSA was dissolved in 5ml 0.1M sodium bicarbonate buffer, the above-mentioned activating drug was added dropwise to the protein solution, stirred at room temperature overnight, dialyzed against PBS at 4°C for 72 hours, and the dialysate was changed 6 times during the period. The dialysate was passed through a filter membrane with a pore size of 0.22 μm under sterile conditions, distributed into ampoule bottles, and stored at -20°C.

[0038] The structural formula of the phenylethanolamine A immune antigen is shown in formula II.

[0039] 2. Synthesis of phenylethanolamine A-coated antigen

[0040] Weigh 10mg of hapten and dissolve in 2ml DMF, add EDC4.3mg, NHS5.1mg, stir at room temperature

[0041] 50mg ...

Embodiment 3

[0044] Example 3, Preparation and Specific Identification of Enzyme-labeled Monoclonal Antibody

[0045] One, the preparation of phenylethanolamine A monoclonal antibody

[0046] 1. Use the immunogen prepared above at 100 μg / mouse, dissolve the immunogen in normal saline and mix it with Freund’s complete adjuvant in equal volumes, inject subcutaneously on the back of the neck and immunize 6-8 week-old Balb / c female mice, after the first immunization On the 7th, 14th, and 28th days, mix the immunogen and Freund's incomplete adjuvant in equal volumes, and each booster immunization once, and 100 μg of the immune complex per mouse 3 days before the fusion, and then booster immunization without Freund's adjuvant.

[0047]2. Proceed according to the conventional method, take the splenocytes of the immunized mice and mix them with the mouse myeloma cells (SP2 / 0) in the logarithmic growth phase, and then slowly add the preheated fusion agent (PEG4000) within 45s for fusion, Suspend e...

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

No PUM Login to View More

Abstract

The invention discloses a phenylethanolamine A hapten, a related artificial antigen and a monoclonal antibody; the invention simultaneously discloses preparation methods and applications of the phenylethanolamine A hapten, the related artificial antigen and the monoclonal antibody. The phenylethanolamine A hapten is the product shown in the formula 1, and the product shown in the formula 1 and a carrier protein can be connected to obtain a phenylethanolamine A antigen. The phenylethanolamine A antigen can be used for preparing a phenylethanolamine A specific antibody. The preparation methods have the advantages of simple and practicable operation, low cost and high yield of the hapten. The phenylethanolamine A artificial antigen can be used for producing the specific antibody aiming at the phenylethanolamine A by immune animals, and can be used for preparing an ELISA kit for detecting phenylethanolamine A residuals; the kit has the advantages of simple and rapid operation, large sample treatment amount, high sensitivity, good specificity, etc.

Description

[0001] technical field [0002] The invention belongs to the technical field of food safety detection, and in particular relates to a preparation method of a phenylethanolamine A hapten, an antigen, a monoclonal antibody and an application thereof. Background technique [0003] β-receptor agonists are used as feed additives in modern farming industry, such as clenbuterol, which is well known to people before. This type of drugs can reduce fat, increase lean meat, and improve the unit economic value of pigs. But they all have dangerous side effects, ranging from irregular heartbeats to heart attacks. [0004] Phenylethanolamine A, also known as "Clonpamine", chemical formula: 2-[4-(4-nitrophenyl) butyl-2-ylamino]-1-methoxyphenethyl alcohol, molecular formula: C19H24N2O4, molecular weight: 344, is a new generation of clenbuterol. During the routine monitoring of illegal drugs in a pig farm at the Zaoshan Town Animal Husbandry and Veterinary Station in Guang'an District, Guan...

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): C07C229/18C07C227/08C07K14/765C07K14/77C07K1/107C07K16/44G01N33/543
Inventor 江海洋温凯吴小平秦誉王照鹏王世恩张桂亮杨艳红金兴王文珺李向梅韩京朋王立淼
Owner 北京维德维康生物技术有限公司
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