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

Group-I type-4 adenovirus long-acting egg yolk antibody preparation method

A technology of egg yolk antibody and adenovirus, which is applied to the preparation methods of peptides, viruses/phages, and botanical equipment and methods, etc., can solve the problem that the immune protection time of egg yolk antibody is short, and the effective immune protection means of avian group I type 4 adenovirus is lacking. and other problems to achieve the effect of solving poor results and avoiding excessive use

Inactive Publication Date: 2018-04-06
TIANJIN RINGPU BIO TECH
View PDF6 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims at the technical defects of the prior art, and provides a method for preparing a long-acting egg yolk antibody against group I adenovirus type 4, so as to solve the technical problem that the prior art lacks effective means of immune protection against poultry group I type 4 adenovirus
[0005] Another technical problem to be solved by the present invention is that conventional egg yolk antibody immune protection time is relatively short in the prior art

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
  • Group-I type-4 adenovirus long-acting egg yolk antibody preparation method
  • Group-I type-4 adenovirus long-acting egg yolk antibody preparation method
  • Group-I type-4 adenovirus long-acting egg yolk antibody preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 (preparation of egg yolk antibody IgY of group I adenovirus type 4)

[0034] Preparation of high-immune eggs: immunize laying hens with group I adenovirus type 4 inactivated vaccine, the immunization dose is 2ml / hens, and the immunization interval is 21 days. After three immunizations, eggs are collected for yolk antibody titer detection. Eggs can be collected with a titer higher than 1:64.

[0035] Extraction and preparation of egg yolk antibody IgY: Soak and sterilize the egg yolk in 1% bromogeramine solution for 15 minutes, dry it for later use; mechanically beat the egg, separate and collect the egg yolk; emulsify with a colloid mill, add acetate buffer at 1:2.5 solution (pH5.2), stirred for 30min, filtered with a plate and frame filter; added 4% caprylic acid to the filtrate, stirred vigorously for 60min, and filtered with polypropylene 750B filter cloth; the filtrate was sterilized by 0.22μm microporous filtration .

Embodiment 2

[0036] Example 2 (preparation of the hexon protein antigen of group I adenovirus type 4)

[0037] Using the co-conserved fragment of the hexon gene of adenovirus type 4 in GenBank, use the software Primer5.0 to design upstream and downstream primers (upstream primer F: 5'-GCGAATTCATGGCGGCCCTCACGCC-3'; downstream primer R: 5'-GCCTCGAGTTACACGGCGTTGCCT-3') ;Using group I type 4 adenovirus as a template, denaturation at 94°C for 45s, annealing at 55°C for 45s, and extension at 72°C for 90s, for a total of 30 cycles of PCR amplification; the PCR product was directly cloned into the pMD-18T vector to construct pMD -18T-FAV; Sequence determination was carried out by the dideoxy chain termination method, pMD-18T-FAV and pET-30a were digested with EcoRI and XhoI, and the digested products were subjected to 1% agarose gel electrophoresis, and the target fragment and pET-30a, then ligated overnight at 16°C; transformed the ligated product into E.coli DH5α competent cells, picked a single...

Embodiment 3

[0038] Example 3 (Titer determination of group I group 4 adenovirus hexon protein antigen and I group 4 type adenovirus egg yolk antibody)

[0039] The potency of group I adenovirus type 4 adenovirus hexon protein antigen and group I group 4 adenovirus type 4 egg yolk antibody was determined by agar diffusion test. The specific operation is as follows: prepare an agar plate containing 8% NaCl and 1% agarose; use plum blossom holes to punch holes on the agar plate;

[0040] Detect the titer of hexon protein antigen: inject the hexon protein antigen into the peripheral holes of the plum blossom holes after doubling dilution, and add the standard positive serum of group I adenovirus type 4 in the center hole, and the sample volume should be filled without overflow. Put the agar plate into a wet box and act at 37°C for 24h to 48h. The maximum dilution factor at which precipitation lines appear in the central well and the peripheral well is the antigen titer of the hexon protein. ...

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 provides a group-I type-4 adenovirus long-acting egg yolk antibody preparation method. According to the technical scheme, on one hand, a group-I type-4 adenovirus inactivated vaccine isutilized for preparing an egg yolk antibody, on the other hand, a group-I type-4 adenovirus hexon protein antigen expression vector is built, and a prokaryotic expression system is utilized for preparing an antigen protein; on the basis, the influence of antigen-antibody valence matching on an antigen-antibody mixed liquor immunoprotection effect is investigated, an optimal compound method is determined on the basis, and a freeze-drying dosage form is designed. According to the method, on the basis of an original adenovirus egg yolk antibody, an adenovirus hexon protein antigen is added, and acompound is prepared. The adenovirus antigen-antibody compound can resist persistent infection caused by a pathogenic microorganism, the overdose of antibiotics is avoided, and the used antigen is not a complete viral antigen but an antigen prepared from a hexon protein of an adenovirus, so that the danger of outward dispersing toxicity can be reduced, and more safety and effectiveness are realized.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a method for preparing a long-acting egg yolk antibody against group I group 4 adenovirus. Background technique [0002] Chicken pericardial effusion and hepatitis syndrome is an acute infectious disease of chickens caused by poultry group I adenovirus type 4. The disease mainly occurs in broiler chickens aged 3 to 6 weeks, and can also occasionally occur in laying hens within 20 weeks of age Or breeder chickens, there is no sex difference, and it rarely occurs in older broilers. Various strains of broiler chickens are susceptible to the disease, and other poultry such as turkeys, geese, and pheasants have also been reported. Clinically, it is characterized by sudden increase in mortality, pericardial effusion and hepatitis syndrome. The infection route of this disease is mainly the respiratory tract, digestive tract and conjunctiva, etc., and can be rapi...

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): C07K16/08C07K16/02C07K14/075C07K1/22C12N15/34C12N15/70
CPCC07K14/005C07K16/02C07K16/081C07K2317/23C12N15/70C12N2710/10022
Inventor 杨燚李守军刘冠星郁宏伟苏建东付旭彬
Owner TIANJIN RINGPU BIO TECH
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