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Epitope vaccine of hog cholera virus and its preparing process

An epitope vaccine, swine fever virus technology, applied in antiviral agents, pharmaceutical formulations, medical preparations containing active ingredients, etc. Sexuality, concentration and intensity of humoral responses, preventing viral infection

Inactive Publication Date: 2008-04-02
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with traditional live attenuated vaccines, the vaccine has the advantages of "marker vaccines", but at the same time has a disadvantage in terms of protection efficacy and production costs

Method used

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  • Epitope vaccine of hog cholera virus and its preparing process
  • Epitope vaccine of hog cholera virus and its preparing process
  • Epitope vaccine of hog cholera virus and its preparing process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, expression and purification of the swine fever virus epitope vaccine of the present invention and its antigenicity detection

[0032] 1. Expression and purification of CSFV epitope vaccine

[0033] 1. Construction of vectors capable of expressing classical swine fever virus epitope vaccines

[0034] 1) Acquisition of primers and marker fragments

[0035] A sequence of the CSF virus protective antigen E0 protein genome was used as a marker fragment (named M, sequence 2 in the sequence listing, Figure 1b), and the introduction of the marker sequence was to facilitate the operation of molecular construction. Other sequences can also be selected as the marker sequence. The marker sequence that meets the requirements of this embodiment should meet the conditions that the sequence should exceed 100 bp in size, and there should be no BglII and XhoI sites within the sequence, preferably no BamHI site.

[0036] The shaded region at the 5' end of the sequence show...

Embodiment 2

[0083] Example 2, Screening and identification of highly immunogenic swine fever virus epitope vaccine

[0084] The swine fever virus vaccines GST-E, GST-2E, GST-4E, GST-5E, and GST-6E were immunized with 50 μg / amount of large-eared white rabbits respectively (2 in each vaccine group, denoted as A and B respectively). ), with GST as the control, complete Freund's adjuvant was used for the first immunization, and incomplete Freund's adjuvant was used for the second and third immunizations. After the first immunization, the second immunization was carried out on the 14th day, and the third immunization was carried out on the 28th day; from the date of the first immunization, blood was collected before immunization on the day of the first immunization to prepare pre-immune serum, On the 14th day after the first immunization, blood was collected before the second immunization to prepare the serum after the first immunization; on the 21st day after the first immunization, blood was...

Embodiment 3

[0088] Embodiment 3, the protective effect identification of the swine fever virus vaccine of the present invention

[0089] 1, the test rabbit of immunization GST (GST immunization group), GST-E (GST-E immunization group), GST-6E (GST-6E immunization group) in the implementation example 2 is two weeks after the third immunization, with pig The attenuated rabbit vaccine (Qianyuanhao Biological Co., Ltd.) was used for the challenge / protection test. Each rabbit was inoculated with 100 minimum rabbit body infection dose (100RMID). After the inoculation, the body temperature curve was recorded within 96 hours. Rectal temperature (rectal temperature). Healthy big-eared white rabbits (non-immunized group) that had not been immunized and had not been exposed to CSFV were used as negative controls. The results are shown in Figure 7a. The results showed that the GST-6E-immunized white rabbits had no stereotyped fever in the challenge test, while the two rabbits in the other immunizati...

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Abstract

The invention discloses a swine fever virus epitope vaccine and a method of preparing the vaccine. An amino acid residue sequence of active protein of the vaccine comprises 6 copies of or more than 6 copies of the amino acid residue sequence of the sequence 1. In the swine fever virus epitope vaccine provided by the invention, the active protein selects very conservative neutralizing epitope to remove the surrounding sequences of the neutralizing epitope, and does not comprise any other virus components or other virus protein sequences; GS is regarded as a connecting peptide, which can more focus on displaying the epitope function, so that the body fluid response to the epitope can be more concentrated and intense.

Description

technical field [0001] The invention relates to a swine fever virus epitope vaccine and a preparation method thereof. Background technique [0002] Classical swine fever virus (CSFV) caused by classical swine fever virus (CSFV) is an important viral disease that endangers the pig industry. It is one of the legally reported severe infectious diseases determined by the World Organization for Animal Health. It is one of the epidemic diseases, and its prevention and control work is highly valued by countries all over the world. At present, except some countries such as the European Union adopt non-vaccination stamping-out policy to eradicate swine fever virus, many countries in the world including my country adopt immunophylactic policy. Although many new vaccines are currently being developed, most of them still have a lot of work to do before they can be put into practical use. At present, the most common CSFV vaccines on the market are live attenuated vaccines, including th...

Claims

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Application Information

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IPC IPC(8): A61K39/187A61P31/14
Inventor 陈应华戚昀张冰清
Owner TSINGHUA UNIV
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