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 vacc

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

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[0031] Example 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 swine fever virus epitope vaccine

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

[0034] 1) Acquisition of primers and labeled fragments

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

[0036] The shaded area at the 5'end of the sequen...

Example Embodiment

[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 50μg / head of big-eared white rabbits (2 in each vaccine group, denoted as A and B respectively) ), with GST as a control, complete Freund's adjuvant for the first immunization, and incomplete Freund's adjuvant for the second and third immunizations. After the first immunization, the second immunization will be performed on the 14th day and the third immunization will be performed on the 28th day; counting from the day of the first immunization, blood will be taken before the 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 post-immunization serum, and the blood was collected on the 21st day after the first immunization to prepare the second...

Example Embodiment

[0088] Example 3. Identification of the protective effect of the swine fever virus vaccine of the present invention

[0089] 1. The experimental rabbits immunized with GST (GST immunization group), GST-E (GST-E immunization group), and GST-6E (GST-6E immunization group) in Example 2 were used two weeks after the third immunization. Plague rabbitization attenuated virus vaccine (Qianyuanhao Biological Co., Ltd.) was tested for challenge / protection. Each rabbit was vaccinated with 100 minimum rabbit body infections (100RMID). After vaccination, the temperature curve was measured within 96 hours, and the measured body temperature was the rabbit. Rectal temperature (rectal temperature). Healthy big-eared white rabbits (non-immunized group) that have not been immunized and not exposed to swine fever virus were used as negative controls. The results are shown in Figure 7a. The results show that the GST-6E-immunized big-eared white rabbits had no stereotype fever in the challenge test, w...

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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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