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

Sulfonamide hapten, and preparation method and application thereof

A sulfonamide drug and hapten technology, applied in the field of hapten and its preparation, can solve the problems of complex pretreatment, tedious and time-consuming, and inability to operate on site, and achieve the effect of efficient detection method, simple synthesis method, and low detection cost

Active Publication Date: 2013-09-11
BEIJING KWINBON BIOTECH
View PDF1 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The microbiological method is simple and low in cost, but it is time-consuming to operate, has poor sensitivity and specificity, and has difficulties in both qualitative and quantitative aspects. It cannot meet the needs of modern residue detection, and some detection results even fail to meet the requirements of the maximum residue limit. ; Physicochemical methods are often used as confirmatory methods for the determination of drug residues in animal tissues, such as gas chromatography (GC), high performance liquid chromatography (HPLC), gas-mass spectrometry (GC-MS), liquid-mass spectrometry (HPLC -MS), etc., these methods are sensitive, accurate, specific, and good in separation, and can simultaneously determine multiple drugs, but require expensive instruments, sample pretreatment is complex, cumbersome and time-consuming, and the cost of detection is high, so they cannot be operated on-site , and requires professionals to operate, so its application is limited; in contrast, ELISA has the advantages of good specificity, high sensitivity, easy operation, low detection cost, suitable for screening and detection of batch samples, etc., and requires Less equipment, easy to learn operation, low cost, can better meet the inspection work of my country's livestock and poultry farmers, slaughterhouses, food companies, government functional supervision departments, etc.

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
  • Sulfonamide hapten, and preparation method and application thereof
  • Sulfonamide hapten, and preparation method and application thereof
  • Sulfonamide hapten, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Synthesis and identification of embodiment 1 sulfa drug hapten

[0021] One, the synthesis of sulfonamide drug hapten (synthetic route such as figure 1 )

[0022] a) Add 0.87g of 6-aminonicotinic acid, 20ml of ethanol, and 3.7ml of 12mol / L hydrochloric acid into a 25ml single-mouth bottle, heat to reflux, react for 8h, monitor by TLC, after the reaction is completed, remove the solvent under reduced pressure, dissolve in saturated NaHCO 3 Aqueous solution, extracted with ethyl acetate, anhydrous NaS 2 o 4 Drying, column chromatography (ethyl acetate / petroleum ether, 2 / 1, v / v) purification;

[0023] b) Add 0.89g of the product obtained in a) into a 25ml single-necked bottle, triethylamine (Et 3 N) 1.6ml, CH 2 Cl 2 10ml, after stirring and dissolving, add the catalyst 4-dimethylaminopyridine (DMAP). Slowly drop into 2ml of dichloromethane solution of 4-acetamidobenzenesulfonyl chloride, TLC monitoring, 5h, after the reaction is completed, treat, dry, and purify by ...

Embodiment 2

[0027] Embodiment 2 sulfa drugs antigen

[0028] The sulfonamide drug hapten is coupled with the carrier protein to obtain the sulfa drug antigen.

[0029] 1. Preparation of immunogen - synthesis of sulfonamide drug hapten-bovine serum albumin conjugate

[0030] Take 12mg of sulfonamide drug hapten and dissolve it in 1ml N,N-dimethylformamide (DMF) to obtain solution I; take 15mg carbodiimide (EDC) and dissolve it fully with 0.2ml water and add it to I, Stir at room temperature for 24 hours to obtain solution II; weigh 40 mg of bovine serum albumin (BSA) to fully dissolve it in 3 ml of PBS (pH7.2), slowly add solution II to the protein solution drop by drop, and leave at room temperature Stir at low temperature for 24 hours; dialyze with 0.01mol / L PBS at 4°C for 3 days, and change the dialysate 3 times a day to remove unreacted small molecular substances to obtain the sulfa drug immunogen; aliquot and store at -20°C for later use.

[0031] 2. Preparation of Coating Source—Sy...

Embodiment 3

[0035] Embodiment 3 sulfa drug monoclonal antibody

[0036] 1. Preparation of monoclonal antibodies against sulfa drugs

[0037] Animal immunization: Inject the immunogen into the body of Balb / c mice with an immunization dose of 150 μg / mouse to make it produce polyclonal antibody serum.

[0038] Cell fusion and cloning: After the measurement result of mouse serum was higher, the splenocytes were taken and fused with SP2 / 0 myeloma cells at a ratio of 8:1, and the cell supernatant was measured by indirect competitive ELISA, and the positive wells were screened. Positive wells were cloned by limiting dilution until hybridoma cell lines secreting monoclonal antibodies were obtained.

[0039] Cell cryopreservation and recovery: the monoclonal hybridoma cell line of sulfa drugs was made into 1×10 9 cells / ml for long-term storage in liquid nitrogen. When recovering, take out the cryopreservation tube, put it into a 37°C water bath to thaw quickly, remove the cryopreservation solutio...

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 hapten, and a preparation method and application thereof, particularly a sulfonamide hapten. The invention also discloses a preparation method and application of the hapten. The kit quick detection product established on the basis of the sulfonamide hapten is convenient to use and low in detection cost; and the detection method is efficient, accurate and quick, can be used for simultaneously detecting mass samples, and is suitable for on-site supervision of sulfonamide residues in animal-derived samples and screening of mass samples.

Description

technical field [0001] The invention relates to a hapten and its preparation method and application, in particular to a sulfonamide drug hapten and its preparation method and application. technical background [0002] Sulfonamides (SAs) are a class of chemotherapeutic drugs with the structure of sulfanilamide used for the prevention and treatment of bacterial infectious diseases. Because of its broad-spectrum, stability, economy, ease of use, and the combination of antibacterial synergists, the effect can be increased by dozens of times. Drugs with sub-therapeutic concentrations are often used as feed additives to prevent the occurrence of diseases, improve the conversion rate of feed and promote There are more than a dozen kinds of SAs commonly used in animal growth. However, these drugs have serious side effects, and their long-term existence in the human body will cause many bacteria to develop drug resistance to sulfa drugs, and they have potential carcinogenicity, so t...

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): C07D213/80C07K14/765C07K14/77C07K14/795C07K14/75C07K14/47C07K1/113C07K16/44G01N33/577
Inventor 何方洋孙震李勇朱亮亮杨昌松余厚美冯才茂杨秀贤
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