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Immune adjuvant of foot-and-mouth disease vaccine and application of immune adjuvant

A technology for foot-and-mouth disease vaccine and immune adjuvant is applied in the field of vaccine adjuvant and its application, preparation of immune adjuvant for foot-and-mouth disease vaccine, and can solve the problems of delayed onset time, undetectable and the like

Active Publication Date: 2012-09-12
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is also found that the recombinant 3D protein can induce cellular immune response and provide a partial protective immune response, that is, the symptoms of the immunized animals after the virulent challenge of the immunized pigs are significantly reduced, and the onset time is delayed, but FMDV neutrality cannot be detected in the serum of the immunized animals Antibody

Method used

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  • Immune adjuvant of foot-and-mouth disease vaccine and application of immune adjuvant
  • Immune adjuvant of foot-and-mouth disease vaccine and application of immune adjuvant
  • Immune adjuvant of foot-and-mouth disease vaccine and application of immune adjuvant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The preparation of the different fragments of embodiment 1 foot-and-mouth disease virus 3D protein

[0032] 1. Bioinformatics analysis of 3D protein gene of foot-and-mouth disease virus:

[0033] The non-structural protein 3D of foot-and-mouth disease virus is a viral RNA polymerase, which is highly conserved in seven serotypes of foot-and-mouth disease virus. The full length of the 3D gene is 1410 nucleotides, encoding a protein with a length of 470 amino acids. DNAStar biological software was used to analyze the sequences of 46 representative strains of seven serotypes and compare them with the GenBank database. The results showed that the nucleotide The homology is higher than 90%, and the amino acid homology is higher than 97%, highly conservative. In this study, the 3D sequence of Asia type 1 FMD virus JS / China / 05 strain was used as a reference sequence to design primers to amplify the target gene fragment and express the protein.

[0034] 2. Gene cloning and pro...

Embodiment 2

[0036] The preparation of embodiment 2 recombinant antigen (EoIgG)

[0037] 1. Bioinformatics analysis of O-type foot-and-mouth disease virus VP1 gene:

[0038] The structural protein VP1 of foot-and-mouth disease virus is the dominant antigen of the virus. Whether it is the isolated and purified natural VP1 protein or the recombinant expression product, it can induce the body to produce protective neutralizing antibodies, which is type-specific. The full length of the foot-and-mouth disease virus VP1 gene consists of 639 nucleotides, encoding a protein with 213 amino acids, and its main epitopes are concentrated in the 140-160 amino acid and the 200-213 amino acid segment. The present invention uses DNAStar biological software to carry out sequence analysis to three strains of porcine foot-and-mouth disease virus O type representative strains (O / China / 99, O / Miandian / 98, O / ZK / 93) that China isolates, and determined that the dominant antigenic epitope is The 135-160th amino ac...

Embodiment 3

[0046] Embodiment 3, vaccine preparation and immune efficacy experiment:

[0047] 1. Vaccine preparation

[0048] The purified recombinant antigen prepared in Example 2 and the different fragments (3D1\3D2\3D3) of the purified 3D protein prepared in Example 1 were quantified by the Bio-Rad quantitative kit and then diluted to an appropriate concentration, purified After the 3D protein fragments and recombinant antigens were configured in a ratio of 1:2 (V / V), an equal volume of oil adjuvant ISA206 (France) was added to emulsify into a vaccine preparation, which was packed in 1ml / tube, which contained 200 μg of recombinant antigen, 3D Protein fragments 100 μg.

[0049] 2. Immunity efficacy test:

[0050] Use the prepared vaccine to inoculate guinea pigs (100 μg antigen + 50 μg 3D protein fragments) by intramuscular route with 0.5 ml per head and inoculate the animal pigs (200 μg antigen + 100 μg 3D protein fragments) with 1 ml per head for immunization The experimental resul...

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Abstract

The invention discloses an immune adjuvant of a foot-and-mouth disease vaccine and application of the immune adjuvant, and belongs to the field of biological vaccines. The immune adjuvant is a 3D protein fragment of foot-and-mouth disease viruses. Experiments prove that after the expressed 3D protein fragment is compatible with antigens, animals are immunized, the immunogenicity of multi-epitope antigens can be enhanced, and high-level protective neutralizing antibodies are produced. In vitro stimulation shows that after the 3D protein fragment of the foot-and-mouth disease viruses is separately adopted or compatible with the multi-epitope antigens, lymphocyte proliferation response can be produced, namely the immune adjuvant can be used for enhancing immune response, distinguishing infection and immunizing the animals and is a good immune stimulant and a vaccine molecular marker.

Description

technical field [0001] The invention relates to a vaccine adjuvant and its application, in particular to an immune adjuvant for preparing foot-and-mouth disease vaccine and its application. It belongs to the field of biological vaccines. Background technique [0002] As a major animal disease, foot-and-mouth disease seriously threatens the healthy and sustainable development of animal husbandry, and also affects animal food safety and foreign trade exports. Once the disease occurs, the loss will be huge and the impact will be bad. Inactivated vaccines, as the main prevention and control materials, play a very important role in the control of FMD epidemics. However, the production of such vaccines requires the construction of high-level biosafety production workshops to prevent pathogens from escaping, and to distinguish between vaccine immunity and natural infection of animals. What's more serious is that incomplete virus inactivation has the risk of causing the epidemic o...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K39/135A61P31/14C07K14/00C12N15/11C12N15/63C12N1/15C12N1/19C12N1/21
Inventor 邵军军常惠芸丛国正林彤高闪电独军政
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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