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

Monoclonal antibody having PEDV neutralizing activity

A monoclonal antibody and active technology, applied in the field of genetic engineering, can solve the problems of poor immune effect and large side effects, and achieve the effects of convenient operation, simple preparation and increasing antibody content.

Active Publication Date: 2019-12-13
CHINA AGRI UNIV
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many problems in the application of PED vaccines at home and abroad, such as poor immune effect and relatively large side effects.

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
  • Monoclonal antibody having PEDV neutralizing activity
  • Monoclonal antibody having PEDV neutralizing activity
  • Monoclonal antibody having PEDV neutralizing activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1, the preparation of the hybridoma cell line that can secrete the monoclonal antibody with PEDV neutralizing activity

[0062]Using PEDV S protein (product of Kaioudi Co., Ltd.) as the immunogen, Freund’s adjuvant and PEDV S protein were fully mixed to form an oil emulsion, and 4 SPF grade 6-8 week-old Balb / c mice were selected for immunization, including Primary immunization and three booster immunizations. Orbital blood was collected 7 days after the second booster immunization, and the antibody titer in the serum was detected by ELISA method, and the mouse with the highest antibody titer and positive value was selected for the last booster immunization, and the spleen cells of the mice were collected 4 days later Hybridoma cell fusion was performed with SP2 / 0 cells by PEG2000. When the hybridoma cells form clonal colonies, the cell supernatant is detected by ELISA method, and the positive wells secreting anti-PEDV antibodies are screened out, and the cor...

Embodiment 2

[0115] Example 2, In vivo experimental verification of the supernatant of a hybridoma cell line secreting a monoclonal antibody with PEDV neutralizing activity

[0116]A large-scale pig farm located in Ganzhou City, Jiangxi Province had a full-scale outbreak of PEDV virus, and newborn piglets had symptoms such as severe vomiting, watery diarrhea, and dehydration. And died 24-72 hours after the onset. After collecting the small intestine samples of sick pigs and carrying out PCR amplification, it was confirmed that the piglets in this pig farm were infected with PEDV virus.

[0117] 1. Detection method of disease material

[0118] 1. RNA extraction by Trizol method

[0119] (1) For the small intestine tissue frozen at -80°C, add 1ml Trizol for every 30-50mg of tissue, and use a homogenizer for homogenization.

[0120] (2) Place the homogenized sample at 15-30°C for 5 minutes.

[0121] (3) Add 0.2ml chloroform for every 1ml Trizol used, cover the tube cap, shake vigorously f...

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 monoclonal antibody having PEDV neutralizing activity. The monoclonal antibody having the PEDV neutralizing activity and provided by the invention is secreted by a monoclonalantibody hybridoma cell strain SWS-2 having the PEDV neutralizing activity. The monoclonal antibody hybridoma cell strain SWS-2 having the PEDV neutralizing activity has the registration number of CGMCC No.18312 in China General Microbiological Culture Collection Center. The monoclonal antibody having PEDV neutralizing activity is prepared, so that newborn piglets orally take the antibody, the infection probability of the piglets can be effectively reduced, and the protection rate reaches 100% in a cohabitant infection experiment. Results show that the monoclonal antibody having PEDV neutralizing activity can effectively increase the antibody content in the newborn piglets and reduce the risk of illness; the method is simple to prepare and convenient to operate and has a great applicationprospect.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a monoclonal antibody with PEDV neutralizing activity. Background technique [0002] Porcine epidemic diarrhea (PED) is a highly contagious porcine intestinal infectious disease with a mortality rate of up to 100% in suckling piglets. The main symptoms are vomiting, watery diarrhoea, dehydration and high mortality of piglets. There is a large amount of yellow feces around the anus of the sick pig, and the necropsy shows that the small intestine is dilated and filled with yellow liquid, the mesentery is congested, the lymph nodes are edema, and the villi are short. Morbidity and mortality are high in lactating piglets, causing significant economic losses. [0003] Porcine epidemic diarrhea virus (PEDV) belongs to coronavirus, and its genome structure is single-stranded positive-strand RNA. The S protein is located on the outer surface of the virion, which is the key to the vi...

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): C12N5/20C07K16/10G01N33/577G01N33/569G01N33/68A61K39/395A61P31/14C12R1/91
CPCA61K2039/505A61P31/14C07K16/10C07K2317/76G01N33/56983G01N33/577G01N33/68G01N2333/17
Inventor 李秋艳史文姝赵兴波
Owner CHINA AGRI UNIV
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