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

Monoclonal antibody specifically combined with porcine rotavirus and application thereof

A monoclonal antibody and porcine rotavirus technology, applied in the direction of antibodies, antiviral agents, antiviral immunoglobulin, etc., can solve the problems of expensive instruments, high background, and time-consuming separation and identification, and achieve rapid detection and high sensitivity Effect

Active Publication Date: 2021-06-01
LUOYANG PULIKE WANTAI BIOTECH
View PDF6 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the methods for detecting porcine rotavirus include virus isolation and identification, serum examination and PCR method, but the isolation and identification, serum examination take too long, and the PCR method is sensitive but requires expensive instruments and professional operators
Although there are commercially available porcine rotavirus colloidal gold test strips, they can only detect group A rotaviruses but not other groups of rotaviruses, and the detection of complex clinical samples will produce a high background leading to Difficult to observe, poor sensitivity and specificity

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 specifically combined with porcine rotavirus and application thereof
  • Monoclonal antibody specifically combined with porcine rotavirus and application thereof
  • Monoclonal antibody specifically combined with porcine rotavirus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Preparation, purification and identification of porcine rotavirus monoclonal antibody

[0061] 1.1 Preparation of monoclonal antibody

[0062] Cultivate the porcine rotavirus OSU strain (purchased from ATCC), centrifuge the harvested virus liquid at 5000 rpm for 30 minutes, get the supernatant at 32000 rpm for 2 hours, and resuspend the precipitate with PBS; prepare 55%, 65 %, 75% density sucrose solution, add it to a centrifuge tube to make a 55%-75% sucrose concentration gradient, add the precipitate suspension, centrifuge at 4°C, 32000 rpm for 2 hours, and harvest a 60%-70% concentration gradient After purification, the protein concentration was determined to be 0.98mg / ml; it was emulsified with Freund's adjuvant, and the final content was 30μg / mouse to immunize mice every 2 weeks. After immunization for 3 times, blood was collected from mice and used Indirect Immunofluorescence (IFA) (inoculate cells with PRoV until 30% of the lesions appear in the cells,...

Embodiment 2

[0104] Preparation and application of embodiment 2 kit

[0105] 2.1 Pairing studies of monoclonal antibodies

[0106] Colloidal gold detection test strips were prepared by using monoclonal antibodies 8C3D6 and 6D4A8 as gold-labeled antibodies and immobilized antibodies, respectively. Dilute the PRoV OSU strain and HN03 strain to 10 5.0 TCID 50 / ml, detection of PRoV OSU strain, HN03 strain and clinically negative feces identified by RT-PCR, the results are shown in Table 5: when using gold-labeled antibody 6D4A8 and immobilized antibody 8C3D6, the dilutions of PRoV OSU strain and HN03 strain were all positive, clinical Negative stools are weakly positive and false positive. Therefore, the gold-labeled antibody 6D4A8 and the immobilized antibody 8C3D6 were selected as the matching method for kit preparation.

[0107] Table 5 Matching results of monoclonal antibodies

[0108] immobilized antibody Gold labeled antibody OSU strain HN03 strain clinically nega...

Embodiment 3

[0154] Example 3 Effect Evaluation of Monoclonal Antibody 8C3D6 in Preventing and / or Treating Virus Infection

[0155] 3.1 Evaluation of the effect of monoclonal antibody 8C3D6 in preventing viral infection

[0156] Screen PEDV, TGEV, PRoV antigen, antibody double-negative 2-3 day old piglets 15, be divided into 3 groups at random, the monoclonal antibody 8C3D6 2mL that the first group and the second group are prepared orally in the embodiment 1 respectively, the third 2 mL of PBS buffer solution was taken orally in the first group, and 24 hours after the oral administration, the first group and the third group were all orally administered 2 mL of PRoV epidemic strain HN03 strain small intestine virus (containing 1000 minimum incidence doses), and the second group was orally administered OSU strain small intestine virus (containing 1000 Minimum incidence dose), observe the clinical symptoms of piglets every day, and observe continuously for 14 days, and evaluate the prevention...

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
Sensitivityaaaaaaaaaa
Sensitivityaaaaaaaaaa
Login to View More

Abstract

The invention provides a variable region sequence of a mouse monoclonal antibody specifically combined with porcine rotavirus, an antibody or an antibody fragment prepared from the variable region sequence, an application of the antibody, and a kit and a pharmaceutical composition prepared from the antibody. The antibody disclosed by the invention can be used for indirect immunofluorescence, immunohistochemistry and the like; the kit disclosed by the invention can be used for effectively detecting the porcine rotavirus, and is high in sensitivity and rapid in detection; and the pharmaceutical composition disclosed by the invention can be used for effectively preventing and treating porcine rotavirus infection.

Description

technical field [0001] The invention relates to a pair of porcine rotavirus monoclonal antibodies, a pharmaceutical composition prepared by using the monoclonal antibodies, a kit and applications thereof, and the invention belongs to the field of biomedicine. Background technique [0002] Porcine rotavirus disease is an acute intestinal infectious disease of pigs caused by porcine rotavirus (PRoV). The main symptoms are anorexia, vomiting and diarrhea. The incidence rate of suckling piglets is high, the incidence rate is generally 50% to 80%, and the mortality rate is generally within 10%. Porcine rotavirus is a virus that can infect both humans and animals. It has a high infection rate in pigs. After infection, the immunity of pigs, especially piglets, will drop sharply, which will induce mixed infections of various pathogens and lead to an increase in the mortality of piglets. or growth retardation. Porcine rotaviruses are divided into 4 subgroups: A, B, C, and E. The ge...

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): C07K16/10C12N5/20A61K39/42A61P31/14G01N33/577G01N33/569G01N33/558
CPCC07K16/10A61P31/14G01N33/577G01N33/56983G01N33/558C07K2317/56C07K2317/92C07K2317/76C07K2317/33A61K2039/505G01N2469/10G01N2333/14
Inventor 田克恭张云静
Owner LUOYANG PULIKE WANTAI 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