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

Polypeptide for inhibiting hog cholera virus infection activity and application thereof

A swine fever virus and bovine viral diarrhea technology, applied in the field of polypeptide design and anti-viral infection research, can solve the problems of irregular feeding conditions, uneven vaccine quality, and unscientific immunization procedures of pigs, and achieve no immunogen. , easy to synthesize and modify, good inhibitory effect

Active Publication Date: 2018-09-28
HENAN ACAD OF AGRI SCI
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease has been prevalent in my country, and it is mainly sporadic in regions; although major farms have paid great attention to CSF, due to unscientific immunization procedures, uneven vaccine quality and non-standard feeding conditions of pigs And so on, resulting in the current CSF is still a serious threat to my country's pig industry

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
  • Polypeptide for inhibiting hog cholera virus infection activity and application thereof
  • Polypeptide for inhibiting hog cholera virus infection activity and application thereof
  • Polypeptide for inhibiting hog cholera virus infection activity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1. Molecular docking and screening of virtual peptide library

[0023] 1. Preparation of E2 protein

[0024] The crystal structure (4JNT) of Bovine Epidemic Diarrhea Virus E2 protein was searched from the PDB database, and the crystal structure was analyzed by a computer program, and the 800th to 900th amino acid residues were selected as the docking setting region for molecular docking.

[0025] 2. Design of virtual peptide library

[0026] The spatial structure of different amino acid residues is established, and the input target polypeptide can be generated in batches with the help of computer programs, so as to meet the automatic calling and processing of molecular docking computer programs. The virtual peptide libraries are all generated in straight chain form without any modification of side chains and head-to-tail amino and carboxyl groups. The generation of a single virtual polypeptide library is preferably no more than four amino acid residues.

[00...

Embodiment 2

[0029] Embodiment 2, P 8 Affinity identification (SPR) with artificially expressed E2 protein

[0030] 1. Dilute the purified CSFV E2 protein with PBS buffer to 1 μg / mL (protein amount), and use EDC / NHS active ester method to separate 1-(3-dimethylaminopropyl)-3-ethyl carbon Diimide / N-hydroxysuccinimide (EDC / NHS) and CSFVE2 protein were injected into the SPR sensor equipped with the amino chip to ensure that EDC / NHS and E2 proteins interacted with the amino chip for 5 minutes, respectively. Coupling of the chip. After the coupling is completed, the sensor can be used to measure the relationship between CSFVE2 protein and P 8 interaction between.

[0031] 2. Inject 250 μL of PBS buffer (pH 7.4) into the sensor, run the PBS buffer at the maximum flow rate (150 μL / min) to reach the signal baseline, and reduce the flow rate of the PBS buffer to 20 μL / min to obtain a relatively stable baseline .

[0032] 3. The synthesized P 8 Dilute with PBS buffer to different concentration...

Embodiment 3

[0034] Embodiment 3, P 8 ELISA identification with artificially expressed E2 protein

[0035] 1. The artificially expressed and purified CSFV E2 protein was coated with ELISA plate at 1 μg / mL (protein amount); in the same way, the purified protein of different viruses, namely bovine viral diarrhea virus E2 protein (BVDV-E2) , Japanese encephalitis virus E2 protein (JEV-E2), circovirus Cap protein (PCV-Cap), pseudorabies virus gE protein (PRV-gD), and 2% bovine serum albumin (BSA), PBS buffer The solution was coated with ELISA plate as a control. Among them, the coated antigens were diluted with carbonate (CBS) buffer, 50 μL per well was added to a 96-well microtiter plate, placed at 4°C overnight, washed with PBST buffer for 5 times, and then the mass fraction 2% BSA solution for blocking.

[0036] 2. Synthesized and biotinylated P at the amino terminal 8The dry powder was diluted with PBS buffer (pH 7.4) to a concentration of 500 ng / mL, and added to the above-mentioned EL...

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 polypeptide for inhibiting hog cholera virus infection activity and application thereof. The sequence of the polypeptide is KRWWSHK (P8). The bovine viral diarrhea virus E2 protein crystal structure is used as a template; homology modeling is performed to obtain a three-dimensional structure of the hog cholera virus E2 protein; a virtual molecular butt joint technology isused for finally obtaining a polypeptide sequence with the high score value, i.e., P8. The artificially synthesized P8 sequences uses ELISA and SPR test for identifying the affinity and specificity ofthe mutual effect with the E2 protein. The result shows that the P8 and the E2 have higher affinity and specificity for combination; then, the virus infection inhibition activity is verified throughqRT-PCR and IPMA tests; the results show that the P8 sequence has relatively good activity for inhibiting the hog cholera virus infection PK-15 cells. The reliable theoretical basis and new idea are provided for further studying virus receptor and anti-virus medicine design.

Description

technical field [0001] The invention relates to a polypeptide for inhibiting swine fever virus infection activity and its application, in particular to the design of a polypeptide targeting swine fever virus E2 protein and research on its anti-swine fever virus infection activity, belonging to the field of polypeptide design and anti-virus infection research. Background technique [0002] With the rapid development of genomics and proteomics, more and more three-dimensional structures of biological macromolecules have been analyzed. As of January 2017, more than 120,000 crystal structures have been imported into the protein database. Further in-depth research on the homology modeling method based on amino acid sequence analysis, for some biological macromolecules whose structure cannot be analyzed under the current technical level, the three-dimensional structure can also be analyzed by modeling. It greatly expands the scope of protein research based on structural analysis. ...

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): C07K7/06A61K38/08A61P31/14G01N33/569
CPCA61K38/00A61P31/14C07K7/06G01N33/56983G01N2333/183
Inventor 王方雨张改平邓瑞广余秋颖邢广旭郝慧芳孙亚宁赵东柴书军
Owner HENAN ACAD OF AGRI SCI
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