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

Vibrio cholerae flagellin monoclonal antibody and antigen capture ELISA kit

A monoclonal antibody, Vibrio cholerae technology, applied in the field of immunology, can solve the problems of non-species specificity and unavoidable cross-reaction of Vibrio species, and achieve good specificity and stability

Active Publication Date: 2013-03-27
BEIJING ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECH CENT
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the previous preparation process of Vibrio cholerae monoclonal antibodies, inactivated VC cells were often used as antigens, and most of the proteins in the cells were common proteins of the genus, which did not have interspecies specificity, resulting in the harvested monoclonal antibodies Interspecies cross-reactivity within the Vibrio genus is difficult to avoid

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
  • Vibrio cholerae flagellin monoclonal antibody and antigen capture ELISA kit
  • Vibrio cholerae flagellin monoclonal antibody and antigen capture ELISA kit
  • Vibrio cholerae flagellin monoclonal antibody and antigen capture ELISA kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Preparation of Vibrio cholerae Flagellin Monoclonal Antibody

[0019] 1. Cell culture and extraction of flagellin

[0020] Vibrio cholerae (Vibrio cholera) (VC-75, isolated and stored in the laboratory) was inoculated in T1N1 medium (purchased from Beijing Land Bridge Technology Co., Ltd., item number CM168), activated at 36°C for 24 hours; a single colony was picked and inoculated in a medium containing 2% NaCl TSA medium (purchased from Beijing Land Bridge Technology Co., Ltd., product number CM417) was grown for 18-24 h; well-grown colonies were evenly spread on the TSA (2% NaCl) medium with a sterile cotton swab, and cultured at 36°C for 18 h. ±2h. The next day, the bacterial cells were scraped off with a sterile cotton swab and suspended in a pre-cooled 0.15M NaCl solution, kept in an ice bath for 15-30 min, homogenized at 1000 rpm for 45 sec, and immediately placed on ice for 10 min. Centrifuge the homogenate at 10,000 rpm for 10 min at 4°C; take the s...

Embodiment 2

[0068] Example 2 Vibrio cholerae flagellin capture ELISA kit

[0069] 1. Composition of Vibrio cholerae Flagellin Capture ELISA Kit

[0070] The solid phase carrier that has been coated with the polyclonal antibody of Vibrio cholerae flagellin is the microtiter plate, the monoclonal antibody of Vibrio cholerae flagellin labeled with horseradish peroxidase, the substrate reaction solution of the enzyme, positive and negative controls, washing solution and reaction termination solution.

[0071] (1) ELISA plate coated with polyantibody

[0072]Dilute the Vibrio cholerae flagellin polyclonal antibody (prepared as in Example 1) with PBS buffer to a concentration of 10 μg / mL, and coat a 96-well EIA high-efficiency binding microtiter plate at 100 μL / well, overnight at 4°C. After taking it out, wash the plate 3 times with PBST and shake dry. 2% BSA was dissolved in PBS solution as a blocking solution, 100 μL / well was added to the microtiter plate, and blocked at 37°C for 1 hour. ...

Embodiment 3

[0085] Embodiment 3 Vibrio cholerae flagellin capture ELISA kit using method

[0086] 1. Processing of samples to be tested

[0087] Take 1 mL of the enrichment solution of the sample to be tested, centrifuge at 10,000 rpm for 2 minutes, discard the supernatant, wash the pellet twice with PBS buffer, and add a small amount of 200 μL PBS to resuspend the pellet. Boiling water bath for 5min.

[0088] 2. Add control and test samples

[0089] Take 100 μL of the sample to be tested and add to the corresponding enzyme-labeled well, positive control and negative control 100 μL / well, tap the side of the sample plate to ensure that the sample evenly covers the bottom of the well, incubate at 37°C for 1 hour, wash the plate with PBST 3 times, and shake dry .

[0090] 3. Add enzyme-labeled monoclonal antibody

[0091] Add 100 μL / well of enzyme-labeled monoclonal antibody working solution (1:1000 dilution of enzyme-labeled monoclonal antibody in PBS buffer), incubate at 37°C for 1 hou...

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
Login to View More

Abstract

The invention discloses a Vibrio cholerae flagellin monoclonal antibody and an antigen capture ELISA kit. The Vibrio cholerae flagellin monoclonal antibody is secreted by a hybridoma cell line with a preservation number of CGMCC No.6754. The Vibrio cholerae flagellin monoclonal antibody can be used for detection of Vibrio cholerae. The invention also discloses a Vibrio cholerae flagellin protein capture ELISA kit.

Description

technical field [0001] The invention belongs to the field of immunology, and in particular relates to a flagellin monoclonal antibody of Vibrio cholerae, a hybridoma cell line and an antigen capture ELISA kit. Background technique [0002] Cholera is a severe intestinal infectious disease caused by Vibrio cholera (VC). The disease has an acute onset, rapid spread, and wide coverage. It is an international quarantine infectious disease and is listed as one of the Class A infectious diseases in my country. , is one of the main foodborne pathogenic microorganisms, which always threatens and brings harm to human beings. Human beings are the only susceptible persons to Vibrio cholerae under natural circumstances, and they are mainly transmitted orally through contaminated water or food. Under certain conditions, after Vibrio cholerae enters the small intestine, it relies on the movement of flagella to pass through the mucus layer on the surface of the mucosa, adheres to the epith...

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): C07K16/12C12N5/20G01N33/68G01N33/577G01N33/569
CPCY02A50/30
Inventor 曾静张蕾马丹张西萌魏海燕刘莉周熙城
Owner BEIJING ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECH CENT
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