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Sheep aphthovirus Asial type multi-epitope recombinant vaccine and preparation method thereof

A foot-and-mouth disease virus and recombinant vaccine technology, applied to sheep foot-and-mouth disease virus Asia1 type multi-epitope recombinant vaccine and preparation thereof, in the field of animal recombinant vaccine, can solve the problem of short duration, inability to meet epidemic prevention and control, and inability to protect against virus and other problems to achieve the effect of protecting against venomous attacks

Active Publication Date: 2010-10-20
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology

However, the large-scale animal experiments all ended in failure (the protection rate of the virus attack did not reach the national standard), mainly because the small molecule antigens developed could not stimulate the body to produce a protective level of neutralizing antibodies, and some even could produce a certain level of protection. Antibodies, but the duration is short, which cannot meet the needs of epidemic prevention and control
In addition, the developed virus particle-like capsid protein can induce the body to produce high-titer protective antibodies, and immunized animals can resist the challenge of homologous viruses, but the low yield and high cost of the antigen limit the application of the vaccine
Recently, people have increased the immunogenicity of vaccines by increasing T cell epitopes and foreign carrier proteins to enhance antigen presentation capabilities, increase molecular weight, and promote B cell maturation and differentiation, enhance immune response, and increase the level of peripheral blood circulating antibodies and duration, although the immunogenicity of the antigen is obviously improved to some extent, but high levels of exogenous carrier protein antibodies can be produced after multiple immunizations, which instead affects the immune effect of the target antigen; in addition, because the researchers used The T cell epitope is not a universal epitope, that is, it cannot be recognized by the major histocompatibility complex (MHC molecule) of all host animals, so it cannot induce an immune response in all animals
In recent years, the epitope vaccine constructed by coupling the main immune factors such as IFN-γ, IL-2, etc. to the epitope small molecule has become a new breakthrough in the research of epitope vaccine, but the immune effect on this animal is not ideal

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  • Sheep aphthovirus Asial type multi-epitope recombinant vaccine and preparation method thereof
  • Sheep aphthovirus Asial type multi-epitope recombinant vaccine and preparation method thereof
  • Sheep aphthovirus Asial type multi-epitope recombinant vaccine and preparation method thereof

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Embodiment 1

[0022] The preparation of embodiment 1 sheep foot-and-mouth disease virus Asia1 type multi-epitope recombinant vaccine

[0023](1) Design and synthesis of multi-epitope genes for foot-and-mouth disease: The whole genome sequence of Asia1 type foot-and-mouth disease virus JS / 05 strain (GenBank accession number: EF149009.1) was selected for multi-epitope design, that is, the main structural protein gene VP1 was selected. Two immunogenic B cell epitopes (136-160aa, 198-211aa) are properly connected in series, that is, 136-160aa-linker-198-211aa. The linker GGSSGG was added between the sites to minimize the impact between adjacent epitopes to ensure the integrity of the structure and function of the epitopes. To increase the potency of the epitope, the concatemer of the two epitopes was repeated once, ie: 136-160aa-linker-198-211aa-linker-136-160aa-linker-198-211aa. In order to insert the multi-epitope fusion gene into the recombinant expression vector, restriction sites were int...

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Abstract

The invention discloses a sheep aphthovirus Asial type multi-epitope recombinant vaccine and a preparation method thereof. The recombinant vaccine comprises the following ingredients: a protein encoded by aphthovirus multi-epitope genes and carrier protein fusion genes, and an aphthovirus 3D protein. The preparation method comprises the following steps: respectively diluting the protein encoded by the aphthovirus multi-epitope genes and the carrier protein fusion genes and the aphthovirus 3D protein; then, uniformly mixing the diluted proteins; adding auxiliary agents into the mixture; carrying out emulsification; and obtaining the sheep aphthovirus Asia1 type multi-epitope recombinant vaccine. Animal model and animal immune efficiency experiments show that the sheep Asial epitope recombinant vaccine of the invention can generate overall immunoprotection reaction, injected immune sheep can be induced to generate high-level neutralizing antibodies, and can also induce the cellullar immunologic response. The recombinant vaccine of the invention can effectively protect animals from the virulent strain attack of the aphthovirus.

Description

technical field [0001] The invention relates to an animal recombinant vaccine, in particular to a Asia1 multi-epitope recombinant vaccine of sheep foot-and-mouth disease virus and a preparation method thereof, belonging to the field of sheep foot-and-mouth disease virus recombinant vaccine. Background technique [0002] As a major animal disease, foot-and-mouth disease not only seriously threatens the healthy development of animal husbandry, but also causes a series of social problems such as people's worries about food safety. Vaccine immunization is still one of the main means to effectively prevent and control FMD. However, most of the currently used vaccines are inactivated vaccines developed from pathogens. The production of such vaccines not only requires a high-level biosafety production workshop to prevent pathogens from escaping, but also requires a distinction between vaccine immunity and naturally infected animals. In addition, incomplete virus inactivation has t...

Claims

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Application Information

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IPC IPC(8): C12N15/42C07K14/09C12N15/62A61K39/135A61K48/00A61P31/14
Inventor 邵军军常惠芸王景锋丛国正林彤独军政高闪电
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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