Unlock instant, AI-driven research and patent intelligence for your innovation.

Preparing method for chloramphenicol hapten and antigen and application of chloramphenicol hapten and antigen in quantum dot immunofluorescence reagent kit

A technology of chloramphenicol and hapten, which is applied in the preparation of chloramphenicol hapten and antigen, and the application field of quantum dot immunofluorescence kit, which can solve the problem of human body producing drug-resistant bacteria beads, destroying the balance state, allergic reaction, etc. problem, to achieve the effect of simple and feasible preparation method, low cost and high sensitivity

Active Publication Date: 2016-03-02
CHINA AGRI UNIV +1
View PDF7 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chloramphenicol residues in animal-derived foods are ingested by the human body for a long time along the food chain, which can cause various diseases
In mild cases, the balance of normal flora in the human body will be destroyed, and the flora will be out of balance, causing the human body to produce drug-resistant bacteria beads, which will bring adverse effects on the use of antibiotics for future illnesses; people with antibiotic allergies will have allergic reactions, endangering health

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
  • Preparing method for chloramphenicol hapten and antigen and application of chloramphenicol hapten and antigen in quantum dot immunofluorescence reagent kit
  • Preparing method for chloramphenicol hapten and antigen and application of chloramphenicol hapten and antigen in quantum dot immunofluorescence reagent kit
  • Preparing method for chloramphenicol hapten and antigen and application of chloramphenicol hapten and antigen in quantum dot immunofluorescence reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1, the preparation of chloramphenicol hapten

[0034] The preparation method of chloramphenicol hapten, concrete operation steps comprise:

[0035] Put 323mg of chloramphenicol raw material into a 10ml one-mouth bottle, dissolve 5ml of pyridine, cool down to 0-5 degrees in an ice bath, add 230mg of p-carboxybenzenesulfonyl chloride in batches, heat up to the outside after 1 hour, stir and react for 18 hours, TLC detection, After the raw material drug chloramphenicol is completely reacted, the reaction solution is poured into 25ml of ice water, and a white precipitate is precipitated, filtered, washed with water, and dried to obtain 450 mg of the hapten, which is detected by mass.

[0036] The structural formula of the hapten is shown in formula Ⅰ:

[0037]

Embodiment 2

[0038] Embodiment 2, the preparation of chloramphenicol artificial antigen

[0039] 1. Synthesis of chloramphenicol immune antigen (CAP-BSA)

[0040] ①Weigh 50mg of BSA, dissolve it in 3.5ml of CB solution, weigh 50 / 67000*60*507=22.7mg of hapten and dissolve it in 1.5ml of DMF, add NHS50 / 67000*120*115.09=10.3mg, EDC50 / 67000*120* 191.7=17.2mg, activated at room temperature for 2 hours;

[0041] ② Add the activation solution dropwise to the CB solution of BSA in an ice bath, react overnight at 4°C, and dialyze with PBS.

[0042] The structural formula of the immunogen is as follows.

[0043]

[0044] 2. Synthesis of chloramphenicol-coated antigen (CAP-OVA)

[0045] ①Weigh 33.5mg of OVA, dissolve in 3.5ml of CB solution, weigh hapten 33.5 / 45000*60*507=22.5mg and dissolve in 1.5ml of DMF, add NHS33.5 / 45000*120*115.09=10.3mg, EDC33. 5 / 45000*120*191.7=17.17mg, activated at room temperature for 2 hours;

[0046] ② Add the activation solution dropwise to the CB solution of OVA...

Embodiment 3

[0049] Embodiment 3, preparation of chloramphenicol specific antibody

[0050] 1. Preparation of polyclonal antibody against chloramphenicol

[0051] Take the immunogen (CAP-BSA) solution prepared in Example 1 and dilute it with pH 7.4, 0.01M PBS buffer to obtain the immunogen dilution, which is used for the preparation of polyclonal antibodies. New Zealand white rabbits were used as immunized animals.

[0052] The immunization process is as follows:

[0053] First immunization: Mix the immunogen dilution with an equal volume of Freund's complete adjuvant to make an emulsifier, inject it subcutaneously at multiple points on the back of the neck, and the immunization dose is 2.5 mg / cause;

[0054] Booster immunization: 4 weeks after the first immunization, 8 weeks later and 12 weeks later, a booster immunization was carried out each time. The immunogen dilution was mixed and emulsified with an equal volume of Freund's incomplete adjuvant, and injected subcutaneously at multiple...

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
Particle sizeaaaaaaaaaa
Wavelengthaaaaaaaaaa
Emission wavelengthaaaaaaaaaa
Login to View More

Abstract

The invention discloses chloramphenicol hapten, corresponding artificial antigen and a monoclonal antibody marked by a quantum dot and further discloses a preparing method and application of the chloramphenicol hapten, the corresponding artificial antigen and the monoclonal antibody marked by the quantum dot. The chloramphenicol antigen can be obtained when the chloramphenicol hapten and carrier protein are connected. The chloramphenicol antigen can be applied to preparing a chloramphenicol specific antibody. The preparing method is easy and convenient to implement and low in cost, and the yield of the hapten is high. The artificial antigen and the chloramphenicol specific antibody marked by the quantum dot can be used for preparing a quantum dot immunofluorescence reagent kit for detecting chloramphenicol residues and have the advantages of being simple, fast, large in quantity of treated samples, high in sensitivity, high in specificity and the like.

Description

technical field [0001] The invention relates to a hapten, an antigen and a preparation method thereof, in particular to a preparation method of a chloramphenicol hapten and an antigen and their application in a quantum dot immunofluorescence kit. Background technique [0002] Chloramphenicol (Chloramphenicol, CAP) is a cheap and highly effective broad-spectrum antibiotic that has a good inhibitory effect on both Gram-positive and Gram-negative bacteria. It was once widely used in agriculture and animal husbandry. However, the residual chloramphenicol in food of animal origin is ingested by the human body for a long time along the food chain, which can cause various diseases. In mild cases, the balance of normal flora in the human body will be destroyed, and flora imbalance will cause the human body to produce drug-resistant bacteria beads, which will bring adverse effects on the use of antibiotics for future illnesses; people with antibiotic allergies will have allergic reac...

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
IPC IPC(8): G01N33/558
Inventor 江海洋沈建忠吴小平王战辉丁双阳李建成史为民温凯王照鹏王世恩王文珺韩京朋
Owner CHINA AGRI UNIV
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com