CN106928327B - Hog cholera virus ligand epitope polypeptide SE242 and application thereof - Google Patents
Hog cholera virus ligand epitope polypeptide SE242 and application thereof Download PDFInfo
- Publication number
- CN106928327B CN106928327B CN201710213707.4A CN201710213707A CN106928327B CN 106928327 B CN106928327 B CN 106928327B CN 201710213707 A CN201710213707 A CN 201710213707A CN 106928327 B CN106928327 B CN 106928327B
- Authority
- CN
- China
- Prior art keywords
- cells
- polypeptide
- csfv
- binding
- virus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/24011—Flaviviridae
- C12N2770/24311—Pestivirus, e.g. bovine viral diarrhea virus
- C12N2770/24322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/24011—Flaviviridae
- C12N2770/24311—Pestivirus, e.g. bovine viral diarrhea virus
- C12N2770/24334—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/18—Togaviridae; Flaviviridae
- G01N2333/183—Flaviviridae, e.g. pestivirus, mucosal disease virus, bovine viral diarrhoea virus, classical swine fever virus (hog cholera virus) or border disease virus
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Virology (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Tropical Medicine & Parasitology (AREA)
- Mycology (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Genetics & Genomics (AREA)
- Gastroenterology & Hepatology (AREA)
- Food Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a hog cholera virus ligand epitope polypeptide SE242 and application thereof, wherein the amino acid sequence of the polypeptide SE242 is as follows: SAGPVRKTSCTFNYAKTGKNKYYEPRDSYF are provided. The invention designs and synthesizes a short peptide positioned on the SE24 polypeptide to obtain the polypeptide SE 242. The test for binding and blocking polypeptide SE242 and PK-15 cells shows that the mean fluorescence intensity of the polypeptide SE242 bound to PK-15 cells is obviously higher than that of a FITC positive control group, the mean fluorescence intensity of CSFV blocking SE242 bound to PK-15 cells is obviously lower than that of the binding test, the fact that the polypeptide SE242 can be effectively bound to the PK-15 cells is confirmed, the binding of the SE242 and the PK-15 cells can be blocked by CSFV, and the SE242 is determined to be a ligand epitope of CSFV bound target cells. Meanwhile, the average fluorescence intensity of the SE242 combined PK-15 cells is higher than that of SE24, the CSFV blocking effect is obviously higher than that of SE24, and the SE242 specificity is better.
Description
Technical Field
The invention belongs to the technical field of molecular pathology and immunology, and particularly relates to a swine fever virus ligand epitope polypeptide SE242 and application thereof.
Background
Classical Swine Fever (CSF) is an acute, febrile, highly contagious disease caused by infection with Classical Swine Fever Virus (CSFV). Is one of the most serious infectious diseases of pigs, causes huge economic loss to the pig industry, is listed as a fulminant infectious disease by the animal health organization in the world, and is also a type of infectious disease regulated by the Ministry of agriculture of China. CSFV belongs to one of members of Flaviviridae and pestivirus, the genome is linear single-strand positive-strand RNA and can code 12 mature virus proteins, wherein C, Erns, E1 and E2 are structural proteins, and the rest are non-structural proteins. Structural proteins Erns, E1 and E2 play important roles in virus recognition, adsorption to host cells, and virus antigenicity. Therefore, the research on the CSFV structural protein is mainly focused on the three proteins.
Viral ligands generally refer to viral surface molecules that specifically bind to viral receptors, i.e., viral attachment proteins. Viral receptors refer to molecular complexes located on the surface of host cells that recognize and specifically bind viral ligands. The specific binding of the viral ligand and the viral receptor on the surface of the target cell is a key link of viral infection, so that the blocking of the binding of the viral ligand and the viral receptor can effectively inhibit the viral disease infection. In recent years, screening antiviral polypeptide drugs and vaccines by using virus receptors or virus ligands as targets of antiviral drugs has become a hot point of research.
Chinese patent discloses CSFV E2 protein ligand epitope polypeptide and application thereof (CN2011101126535), and the disclosed polypeptide amino acid sequence is
VHASDERLGPMPCRPKEIGSSAGPVRKTSCTFNYAKTGKNKYYEPRDSYF, having a molecular weight of 5.73 kDa; and the TCID of CSFV was determined by infecting and harvesting CSFV using PK-15 cells as target cells50And infection ratio, performing binding test, virus blocking test and polypeptide blocking test of the synthesized polypeptide and target cellsThe CSFV infects the target cell test, screens the ligand epitope polypeptide which can restrain the virus infection and is combined with the target cell, and carries on the preliminary location to the CSFV E2 protein ligand epitope. However, the polypeptide amino acid sequence of the above patent is long, the ligand positioning is not accurate enough, the synthesis cost is too high in the practical application process, and the application and popularization of the polypeptide are greatly limited, so that the accurate positioning of the ligand epitope polypeptide is very important. At present, no report related to the precise positioning of the E2 protein ligand epitope is found.
Disclosure of Invention
In order to solve the problems, the invention aims to provide a swine fever virus ligand epitope polypeptide SE242 and application thereof, wherein the polypeptide can effectively bind to PK-15 cells, the binding of the SE242 and the PK-15 cells can be blocked by CSFV, and the SE242 is determined as a ligand epitope of the CSFV binding target cells.
In order to achieve the purpose, the invention adopts the technical scheme that:
hog cholera virus ligand epitope polypeptide SE242, wherein the amino acid sequence of the polypeptide SE242 is as follows: SAGPVRKTSCTFNYAKTGKNKYYEPRDSYF are provided.
Application of a hog cholera virus ligand epitope polypeptide SE242 in PK-15 cell binding and CFSV blocking SE242 in PK-15 cell binding experiments.
An application of a CSFV ligand epitope polypeptide SE242 in inhibiting the infection of PK-15 cells by CFSV is provided.
An application of a CSFV ligand epitope polypeptide SE242 in the development of a medicament or a vaccine for inhibiting CSFV from infecting a target cell.
The invention has the beneficial effects that:
1. the invention designs and synthesizes a short peptide positioned on the SE24 polypeptide to obtain the polypeptide SE 242. The results of the binding and blocking test of the polypeptide SE242 and PK-15 cells show that the mean fluorescence intensity of the polypeptide SE242 bound to the PK-15 cells is obviously higher than that of a FITC control group, the mean fluorescence intensity of CSFV blocking SE242 bound to the PK-15 cells is obviously lower than that of the binding test, the polypeptide SE242 can be effectively bound to the PK-15 cells, the binding of the SE242 and the PK-15 cells can be blocked by CSFV, and the SE242 is determined to be a ligand epitope of CSFV bound target cells. Meanwhile, the average fluorescence intensity of the SE242 combined PK-15 cells is higher than that of SE24, the CSFV blocking effect is obviously higher than that of SE24, and the specificity of the ligand epitope SE242 is proved to be stronger than that of SE 24.
2. The polypeptide disclosed by the invention has a shorter amino acid sequence, can be used for further accurately positioning the ligand epitope of the hog cholera virus combined target cell, and is greatly reduced in cost in practical application compared with the polypeptide disclosed by the prior art.
3. The combination test and the blocking test prove that after the CSFV is combined with the virus receptor of PK-15 cells, the combination of the polypeptide and the virus receptor is prevented, and the combination of the polypeptide SE242 and the CSFV on the similar cell receptor is proved. The polypeptide is an ideal drug target for preventing and treating swine fever, and lays a foundation for further developing and researching novel antiviral drugs and vaccines. Meanwhile, a new way is provided for preventing and treating the swine fever.
Drawings
FIG. 1 shows the detection of CSFV (200X) propagated in culture by the cytoimmunochemical staining technique. In the figure, A is PK-15 cells infected with CSFV; b is a blank control cell.
FIG. 2 is an LC-MS/MS map of polypeptide SE 242. The abscissa represents the mass-to-charge ratio (m/z) of the ion and the ordinate represents the intensity of the ion current (100% of the maximum ion current intensity).
FIG. 3 shows the results of the binding and blocking assay of polypeptide SE242 to PK-15 cells. In the figure, a is FITC control; b is polypeptide SE242 combined with PK-15 cells; c is the combination of CSFV blocking polypeptide SE242 and PK-15 cells.
Detailed Description
The following examples further illustrate the embodiments of the present invention in detail.
Example 1
The invention carries out multi-index measurement on the used Shimen strain (cytotoxin) of the classical swine fever virus to ensure that the CSFV strain (cytotoxin) of the Shimen strain can meet the use requirement of the invention.
1. ELISA titer determination of CSFV
Infecting cultured single-layer PK-15 cells with hog cholera virus Shimen strain (cytotoxin), culturing at 37 deg.C for 48-56h, harvesting virus, repeatedly freezing and thawing for 3 timesThe crushed cells are disintegrated to obtain virus, the virus is centrifuged at 4 ℃ and 3000rpm for 10min, the precipitate is removed, and the supernatant is the virus propagation solution. The harvested virus fluid was tested with classical swine fever virus antigen detection kit (SERELISA HCV Ag Mono Indirect). The virus liquid was diluted 1:50, 1:100, 1:200, 1:400 times, each dilution was repeated 6 times, and the test results are shown in table 1. To compare with negative control OD450The titer of the virus was found to be 1:200 in the dilution having a value ratio of 2.2 times and a total value of 0.2 or more.
TABLE 1 detection of Swine fever Virus potency
Note: "+" represents positive and "-" represents negative.
2. TCID of CSFV50Measurement of
The infectivity, i.e. virulence, of the virus culture is tested. There are 2 methods for determining virus virulence, one is to determine the median Lethal Dose (LD) using experimental animals50) Or measuring infection amount (ELD) of half of chick embryo on chick embryo50) (ii) a Another method is to determine the amount of infection (TCD) in half of the cell cultures on tissue cells50Also known as TCID50)。TCID50The calculation can be performed by the Reed-Muench method, interpolation method and Karber method. The invention adopts a fixed PK-15 cell dilution virus method to calculate the TCID of the propagated virus50The results are shown in Table 2, at a dilution of 10 in the virus solution-1、10-2、10-3At the time, the inoculated 4-well cells were all infected, and the dilution was 10-4Is, 2 wells infected, dilution 10-5、10-6All cells were not infected, and normal cells were not infected as a blank. TCID was calculated according to the above test results by Reed-Muench's method50。
TABLE 2 calculation of TCID for CSFV by fixed PK-15 cell dilution Virus method50
The log of the dilution of the virus higher than 50% in this test was-3, the distance ratio was 0.5 and the difference between the logs of the dilutions was-1.
LogTCID50＝0.5×(–1)+(–3)＝–3.5
TCID50＝10-3.5Meaning that the virus solution is diluted 103.5In addition, half of the cells were infected by 100. mu.L of virus solution per well.
3. Determination of the multiplicity of infection (MOI) of CSFV
Multiplicity of infection or infection ratio (multiplicity of infection): refers to the ratio of the number of cells infected with the virus to the total number of cells in a system. 2 × TCID50After CSFV infects PK-15 cells, the cultured and propagated CSFV is detected by using a cytoimmunochemical staining technique, and the CSFV infects PK-15 cells to be brownish red (figure 1A) and the normal control PK-15 cells do not develop color (figure 1B). The MOI is calculated by counting the total number of infected cells and cells per well of a 96-well cell culture plate and analyzing the assay results. When counting cells, a drop of cell suspension is taken to be placed on a cell counting plate, the total number of cells in four large squares on an oblique diagonal is calculated under an inverted microscope, and the calculation formula is as follows:
total number of cells per well n/4 × dilution factor × 104X. milliliters of cell fluid (n is the total number of cells in the four large squares).
As a result: the average number of cells infected per well in a 96 well cell culture plate was 150; the total number of the cells in the four large squares of the counting plate is 280, the milliliter number of the cell sap in each hole is 0.1ml, and the dilution ratio is 10-1Fold, the total number of cells per well was found to be 7.0X 103Thus, the MOI of the cultured propagated virus is: 1:47.
4. Conclusion
ELISA Titers and TCID by determination of CSFV50And multiplicity of infection (MOI), confirming that the strain of Shimen strain (cytotoxic) CSFV can meet the needs of the present study.
Example 2 Synthesis of polypeptide
The research shows that the polypeptide SE24 (amino acid sequence: CVHASDERLGPMPCRPKEIGSSAGPVRKTSCTFNYAKTGKNKYYEPRDSYF) can effectively bind to PK-15 cells, can effectively inhibit CSFV from infecting PK-15 cells, and CSFV can block SE24 from binding to PK-15 cells, so that the polypeptide SE24 is proved to be a CSFV E2 protein ligand epitope polypeptide. To further pinpoint ligand epitope information on polypeptide SE24, 1 short peptide on polypeptide SE24 was designed and synthesized, and the LC-MS/MS map of synthetic peptide SE242 is shown in FIG. 2, and the sequence is as follows:
SE242：SAGPVRKTSCTFNYAKTGKNKYYEPRDSYF(SEQ ID NO.1)
example 3 binding and blocking assay of SE242 with PK-15 cells
Collecting well-grown PK15 cells, washing with PBS 3 times, digesting the cells with 1% trypsin for about 1min, discarding the digestion solution, adding DMEM containing 10% fetal calf serum for suspension, centrifuging at 1000rpm for 5min, resuspending PK15 cells with appropriate amount of PBS, counting, and adjusting cell concentration to 1.0 × 106One/ml for standby.
Binding assay of SE242 to PK-15 cells:
the PK15 cell suspension (cell concentration of 1.0X 10) was added6Pieces/ml) are mixed uniformly and put into an EP tube with 100 mul of each tube, namely the cell number reaches 1.0 multiplied by 105And (4) respectively. Adding polypeptide SE242 (labeled by FITC) into the tube to enable the final concentration of SE242 to reach 0.2mg/ml, setting FITC positive control, repeating the steps three times, keeping away from light on ice for 2h, washing with PBS for 3 times, centrifuging at 1000rpm for 5min each time, adding 800 mu l of PBS after the last centrifugation to resuspend cells, detecting by a flow cytometer, counting 10000 cells, and judging the analysis result.
CSFV blocking SE242 binding assay to PK15 cells:
1.0ml of the PK15 cell suspension (cell concentration 1.0X 10) was taken6One/ml), 200. mu.l of TCID was added50Is 10-3.5The CSFV solution is mixed evenly and acted for 40min at 4 ℃. Centrifuging to remove CSFV solution, washing with PBS for 2 times, re-suspending cells with PBS, mixing well, and subpackaging in EP tube to make the number of cells in each tube reach 1.0 × 105And (4) respectively.
Adding polypeptide SE242 (labeled by FITC) into the tube to enable the final concentration of SE242 to reach 0.2mg/ml, setting FITC positive control, repeating the steps three times, keeping away from light on ice for 2h, washing with PBS for 3 times, centrifuging at 1000rpm for 5min each time, adding 800 mu l of PBS after the last centrifugation to resuspend cells, detecting by a flow cytometer, counting 10000 cells, and judging the analysis result.
The binding test and the blocking test show that the mean fluorescence intensity of the polypeptide SE242 bound to PK-15 cells is obviously higher than that of a FITC control group, the mean fluorescence intensity of CSFV blocking SE242 bound to PK-15 cells is obviously lower than that of the binding test (figure 3 and table 3), the polypeptide SE242 can be effectively bound to the PK-15 cells, the binding of the polypeptide SE242 and the PK-15 cells can be blocked by CSFV, and the SE242 is determined as a ligand epitope of CSFV binding target cells. Further, ligand epitope information of the CSFV combined target cell is accurate, and a foundation is laid for developing a medicament or vaccine for inhibiting the CSFV from infecting the target cell.
TABLE 3 results of the polypeptide SE242 binding to PK-15 cells and CSFV-blocking polypeptide binding assay
Note: the data in the same row are marked with different letters to show significant difference (P < 0.05); shoulder marks with the same letter or no letter designation indicate no significant difference (P > 0.05).
TABLE 4 comparison of SE24, SE242 polypeptide binding to PK-15 cells and CSFV blocking polypeptide binding assay results
Note: the data in the same row are marked with different letters to show significant difference (P < 0.05); shoulder marks with the same letter or no letter designation indicate no significant difference (P > 0.05).
From table 4, it can be seen that the fluorescence intensity difference between SE24 and SE242 and PK15 cell binding test is significant, and the fluorescence intensity difference between SE24 and SE242 by CSFV blocking test is significant. SE242 was significantly more effective than SE24 in binding to PK-15 cells and blocking of its binding by CSFV.
Example 4 binding verification of other short peptides
The results of the binding test and the blocking test show that SE242 can effectively bind to PK-15 cells, CSFV can block SE242 from binding to PK-15 cells, and in order to further pinpoint ligand epitope information on SE24 polypeptide, 3 short peptides on SE24 polypeptide are designed and synthesized, and the sequences are as follows:
SE24-1：CVHASDERLGPMPCRPKEIG(SEQ ID NO：2)
SE24-2：PKEIGSSAGPVRKTSCTFNYA(SEQ ID NO：3)
SE24-3：TFNYAKTGKNKYYEPRDSYF(SEQ ID NO：4)
after multiple times of binding tests of the 3 short peptides with PK-15 cells, the three short peptides are proved not to be bound with the PK-15 cells.
According to the invention, after 1 short peptide SE242 is designed and synthesized from the SE24 polypeptide, the short peptide is still combined with a target cell, and the combination test and the virus blocking combination test show that the combination effect of SE242 and the target cell and the combination thereof are blocked by classical swine fever virus is obviously higher than that of SE 24. The polypeptide SE242 of the invention is proved to have good specificity and accuracy. After 3 short peptides are further designed and synthesized from the SE24 polypeptide, the 3 short peptides are not combined with target cells, which indicates that the more accurate positioning of CSFV ligand epitope is to be further screened and researched.
The foregoing description is only a preferred embodiment of the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
SEQUENCE LISTING
<110> institute of science and technology of Henan
<120> hog cholera virus ligand epitope polypeptide SE242 and application thereof
<160>4
<170>PatentIn version 3.5
<210>1
<211>30
<212>PRT
<213> Artificial sequence
<400>1
Ser Ala Gly Pro Val Arg Lys Thr Ser Cys Thr Phe Asn Tyr Ala Lys
1 5 10 15
Thr Gly Lys Asn Lys Tyr Tyr Glu Pro Arg Asp Ser Tyr Phe
20 25 30
<210>2
<211>20
<212>PRT
<213> Artificial sequence
<400>2
Cys Val His Ala Ser Asp Glu Arg Leu Gly Pro Met Pro Cys Arg Pro
1 5 10 15
Lys Glu Ile Gly
20
<210>3
<211>21
<212>PRT
<213> Artificial sequence
<400>3
Pro Lys Glu Ile Gly Ser Ser Ala Gly Pro Val Arg Lys Thr Ser Cys
1 5 10 15
Thr Phe Asn Tyr Ala
20
<210>4
<211>20
<212>PRT
<213> Artificial sequence
<400>4
Thr Phe Asn Tyr Ala Lys Thr Gly Lys Asn Lys Tyr Tyr Glu Pro Arg
1 5 10 15
Asp Ser Tyr Phe
20
Claims (4)
1. The hog cholera virus ligand epitope polypeptide SE242 is characterized in that the amino acid sequence of the polypeptide SE242 is as follows: SAGPVRKTSCTFNYAKTGKNKYYEPRDSYF are provided.
2. The use of the CSFV ligand epitope polypeptide SE242 according to claim 1 for the preparation of a medicament or vaccine for binding PK-15 cells.
3. The use of the CSFV ligand epitope polypeptide SE242 according to claim 1 in the preparation of a medicament or vaccine for inhibiting infection of PK-15 cells by CFSV.
4. The use of the classical swine fever virus ligand epitope polypeptide SE242 according to claim 1, in the preparation of a medicament or vaccine for inhibiting CSFV infection of target cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710213707.4A CN106928327B (en) | 2017-04-01 | 2017-04-01 | Hog cholera virus ligand epitope polypeptide SE242 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710213707.4A CN106928327B (en) | 2017-04-01 | 2017-04-01 | Hog cholera virus ligand epitope polypeptide SE242 and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106928327A CN106928327A (en) | 2017-07-07 |
| CN106928327B true CN106928327B (en) | 2020-02-04 |
Family
ID=59426486
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710213707.4A Expired - Fee Related CN106928327B (en) | 2017-04-01 | 2017-04-01 | Hog cholera virus ligand epitope polypeptide SE242 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106928327B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108084250B (en) * | 2017-11-10 | 2021-07-06 | 浙江美保龙生物技术有限公司 | Small molecular polypeptide for specifically inhibiting hog cholera virus proliferation and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1338310A (en) * | 2000-08-10 | 2002-03-06 | 清华大学 | Epitope vaccine of hog cholera virus and its preparing process |
| CN102268079A (en) * | 2011-05-03 | 2011-12-07 | 新乡学院 | (Classical swine fever virus) CSFV E2 protein ligand epitope peptide and application thereof |
| CN106188250A (en) * | 2016-03-25 | 2016-12-07 | 武汉科前生物股份有限公司 | The epitope simulative peptide of a kind of CSFV E 2 protein and preparation method and application |
| CN106456741A (en) * | 2014-05-23 | 2017-02-22 | 勃林格殷格翰动物保健有限公司 | Recombinant classical swine fever virus (csfv) comprising substitution in the tav epitope of the e2 protein |
-
2017
- 2017-04-01 CN CN201710213707.4A patent/CN106928327B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1338310A (en) * | 2000-08-10 | 2002-03-06 | 清华大学 | Epitope vaccine of hog cholera virus and its preparing process |
| CN102268079A (en) * | 2011-05-03 | 2011-12-07 | 新乡学院 | (Classical swine fever virus) CSFV E2 protein ligand epitope peptide and application thereof |
| CN106456741A (en) * | 2014-05-23 | 2017-02-22 | 勃林格殷格翰动物保健有限公司 | Recombinant classical swine fever virus (csfv) comprising substitution in the tav epitope of the e2 protein |
| CN106188250A (en) * | 2016-03-25 | 2016-12-07 | 武汉科前生物股份有限公司 | The epitope simulative peptide of a kind of CSFV E 2 protein and preparation method and application |
Non-Patent Citations (1)
| Title |
|---|
| 猪瘟病毒受体与配体研究进展;冯春花等;《中国兽医杂志》;20151118;第51卷(第9期);第75-79页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106928327A (en) | 2017-07-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113230395B (en) | Beta coronavirus antigen, beta coronavirus bivalent vaccine, preparation methods and applications thereof | |
| CN111821433B (en) | mRNA vaccine and synthetic method and kit thereof | |
| JP5006782B2 (en) | Vaccines containing attenuated pestiviruses | |
| Sultan et al. | Protective efficacy of different live attenuated infectious bronchitis virus vaccination regimes against challenge with IBV variant-2 circulating in the Middle East | |
| CN109485703B (en) | Foot-and-mouth disease A-type structural protein VP1 antigen epitope polypeptide and application thereof | |
| CN103751774A (en) | Recombinant cell line for stably expressing classical swine fever virus E2 protein, and applications of the same in preparation of subunit vaccines and diagnosis reagents of classical swine fever | |
| CN105331636A (en) | Recombination cell line for stable expression of classical swine fever virus E2 and application thereof | |
| KR20100020037A (en) | Vaccines containing canine parvovirus genetic variants | |
| CN111848786A (en) | Monoclonal antibody, preparation method and application thereof | |
| CN102399757A (en) | Immunogenic compositions and uses thereof | |
| CN113150084B (en) | Nanocrystallized coronavirus antigen and application thereof | |
| CN103751773A (en) | Recombinant BHK cell line for stably expressing classical swine fever virus E0-E1-E2 protein, and applications of the same in preparation of vaccines and diagnosis reagents of classical swine fever | |
| CN106928327B (en) | Hog cholera virus ligand epitope polypeptide SE242 and application thereof | |
| CN106866799B (en) | Hog cholera virus ligand epitope polypeptide SE241 and application thereof | |
| CN102406929B (en) | Co-expressed molecular adjuvant enhanced divalent foot and mouth disease protein engineering vaccine | |
| CN108503696A (en) | A kind of zika virus subunit vaccine of yeast cell to express | |
| CN105974119A (en) | G type enterovirus direct immunofluorescent reagent and kit | |
| CN105254724B (en) | Truncated-type Hepatitis C Virus HCV NS3 antigens and its preparation and application | |
| CN104774249B (en) | Porcine epidemic diarrhea virus M albumen affinity peptide and its screening technique | |
| CN114349829A (en) | Identification of ALV-J MHC-B2 restricted epitope peptide and application thereof | |
| CN110903386B (en) | Fully human monoclonal antibody with high neutralizing activity and resisting chikungunya fever and application | |
| KR20130058706A (en) | Parapoxvirus expressing the vp60 major capsid protein of the rabbit haemorrhagic disease virus | |
| US9315873B2 (en) | Marker vaccine for classical swine fever | |
| CN111675753A (en) | Avian influenza virus epitope screening method and application | |
| CN104193827A (en) | Hepatitis C virus HVR1 (Hypervariable Region 1) fusion antigen and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20200204 Termination date: 20210401 |