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

Fusion protein as well as preparation method, application, expression system and vaccine thereof

A fusion protein and protein technology, applied in the biological field, can solve the problems of vaccination failure, the decline of the body's disease resistance, and the decline of the ability of lymphocyte antibodies.

Inactive Publication Date: 2021-10-22
TECON BIOLOGY CO LTD
View PDF6 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] After PCV infection, the ability of lymphocytes to produce antibodies decreases, the cellular immunity decreases, and the ability of alveolar macrophages to phagocytize and clear pathogens also decreases, and the activity of suppressor T cells increases, resulting in a decrease in the body's disease resistance, thereby making other pathogens more vulnerable. Easy to invade, can be secondary infected with porcine reproductive and respiratory syndrome, mycoplasma, etc., resulting in the failure of various vaccinations

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
  • Fusion protein as well as preparation method, application, expression system and vaccine thereof
  • Fusion protein as well as preparation method, application, expression system and vaccine thereof
  • Fusion protein as well as preparation method, application, expression system and vaccine thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0066] A method for preparing the above fusion protein, comprising expressing the gene encoding the fusion protein in a host.

[0067] The method has the advantages of simple preparation method, low cost, powerful vaccine function and the like.

[0068] The host in the present invention can be, but not limited to, an Escherichia coli expression system, a yeast expression system, an insect expression system, a plant expression system or a mammalian expression system. Since the protein expressed by mammalian cells is translated and processed, its structure and biological properties are closer to natural proteins, so the present invention preferably uses a mammalian expression system to express the gene encoding the fusion protein.

[0069] In a preferred embodiment of the present invention, a mammalian expression system is used to express the gene encoding the fusion protein. Since the protein expressed by mammalian cells is translated and processed, its structure and biologica...

Embodiment 1

[0103] Example 1 Expression, identification and purification of fusion protein PA-C3d-Cap-K

[0104] According to NCBI report Pseudomonas exotoxin A (Pseudomonas aeruginosaPA, accession number: CP007224.1, see SEQ ID NO.1) gene sequence, porcine complement C3d (see SEQ ID NO.8) gene sequence, PCV2-ADDLPP 10069 strain (accession number: EU594437.1) Cap protein gene sequence (see SEQ ID NO.2) and PCV3-US / MN strain (NCBI accession number is: KX898030.1) Cap protein gene sequence (see SEQ ID NO. 3) Analyze and finally design Pseudomonas exotoxin A (PEA) domain I and II proteins, porcine complement C3d, porcine circovirus Cap protein dominant antigenic epitope and carboxyl terminal part containing SEQ ID NO.4 polypeptide Select flexible amino acids (SEQ ID NO.7) and connect them in series in a certain combination to express a fusion protein with good immunogenicity.

[0105] 1.1 Construction and identification of PEA domain I and II and Cap gene cloning vector

[0106] According ...

Embodiment 2

[0129] The preparation of embodiment 2 porcine circovirus vaccine

[0130] Dilute the fusion protein prepared in Example 1 with PBS solution, mix the diluted fusion protein solution with SEPPICISA201R VG adjuvant at a mass fraction of 50%, stir at a speed of 8000r / min for 10min, and add 0.01% (volume ratio) thimerosal solution, so that the final concentration does not exceed 1 / 10,000. After fully oscillating and mixing, according to the requirements of the appendix of the current version of the Chinese Veterinary Pharmacopoeia, after passing the sterility test, viscosity measurement, and stability measurement, place it in 4 ℃ for later use.

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
concentrationaaaaaaaaaa
Login to View More

Abstract

The invention provides a fusion protein as well as a preparation method, application, an expression system and a vaccine thereof, and relates to the technical field of biology. The fusion protein provided by the invention comprises a pseudomonas exotoxin A structural domain segment, a porcine circovirus type 2 Cap protein dominant antigen epitope segment, a porcine circovirus type 3 Cap protein dominant antigen epitope segment and a carboxyl terminal part. According to the invention, dominant epitopes of PCV2 and PCV3Cap proteins are screened, the dominant epitopes are good in antigenicity and easy to highly express, then the dominant epitopes are fused with part of functional fragments of PEA, and meanwhile, carboxyl terminal sequences are added to the C terminal of the fusion protein and the immunogenicity is enhanced, so that the solubility of the fusion protein is improved. The obtained fusion protein has the advantages of targeting positioning, good antigenicity and high expression quantity. The vaccine prepared by taking the fusion protein as an antigen has the advantages of good safety, stable titer, no toxic or side effect and capability of activating cellular immune response.

Description

[0001] This application is a divisional application with an application date of September 19, 2018, an application number of 201811104817.8, and an invention title of "Fusion Protein and Its Preparation Method, Application, Expression System and Vaccine". technical field [0002] The invention relates to the field of biotechnology, in particular to a fusion protein and its preparation method, application, expression system and vaccine. Background technique [0003] Porcine circovirus (Porcine circovirus, PCV) belongs to Circoviridae, single-stranded, circular DNA virus. It is mainly divided into non-pathogenic porcine circovirus type (PCV1) and pathogenic porcine circovirus type (PCV2). It is currently known that the Cap protein is a structural protein of PCV and has good immunogenicity. In October 2016, R. Palinski of Kansas State University in the United States and T.G.Phan of the University of California, San Francisco reported a new PCV genotype, called PCV3. The virus...

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): C07K19/00C07K16/08C12N15/62C12N15/85G01N33/569A61K39/12A61K39/295A61P31/20
CPCA61K39/12A61P31/20G01N33/56983C12N15/85C07K14/005C07K14/21C07K16/081A61K2039/552A61K2039/70C07K2319/55C12N2750/10034C12N2750/10022
Inventor 贺笋张伟李俊辉李延涛程兰玲高窦潘晓梅徐龙飞师小潇
Owner TECON BIOLOGY CO LTD
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