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A-type antigen polypeptide, fusion antigen polypeptide and vaccine of foot and mouth disease virus

A technology of foot-and-mouth disease virus and antigenic polypeptides, applied in the directions of virus antigenic components, virus peptides, multivalent vaccines, etc., can solve the problem of inability to A-type strain antigens, fusion polypeptides and vaccine work enlightenment, no cross-protection effect, mutual immunity, etc. question

Active Publication Date: 2016-03-23
SHANGHAI SHEN LIAN BIOMEDICAL CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the previous Chinese patent CN102180952A (its application date is May 10, 2011, and the invention name is "Foot-and-mouth disease virus type A antigen polypeptide, fusion antigen polypeptide and vaccine"), although it discloses the design for O / MYA98 / BY / 2010 strain O-type antigen polypeptides, fusion antigen polypeptides and vaccines, but this technical solution is only applicable to the O / MYA98 / BY / 2010 strain itself, and the diversity of FMDV serotypes has caused the vaccine to be used among animals infected by different serotypes. There is no protective effect between them, and there is no cross-protective effect between the O-type and A-type strains of foot-and-mouth disease virus. Therefore, the disclosure of the O-type antigen polypeptide, the fusion antigen polypeptide and the vaccine cannot bring any impact on the development of the A-type strain antigen, fusion polypeptide and vaccine. work inspiration
[0007] The high variability of foot-and-mouth disease virus and the mutual immunity between the various serotypes usually bring great difficulties to the prevention and control of foot-and-mouth disease

Method used

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  • A-type antigen polypeptide, fusion antigen polypeptide and vaccine of foot and mouth disease virus
  • A-type antigen polypeptide, fusion antigen polypeptide and vaccine of foot and mouth disease virus
  • A-type antigen polypeptide, fusion antigen polypeptide and vaccine of foot and mouth disease virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0158] Embodiment 1. The solid-phase synthesis of polypeptide

[0159] Equipment: PSI300B automatic peptide synthesizer, using RinkAmideMBHA resin as a solid phase carrier; PallCentrasette tangential flow ultrafiltration system, using Omega series membrane cassettes.

[0160] Raw materials and reagents: 9-fluorenylmethoxycarbonyl (Fmoc)-protected amino acid is used as the starting material, and other reagents used in the solid-phase synthesis are detailed in the following parts of the examples. All the raw materials and reagents used in the synthesis can be purchased in the market or obtained after simple preparation by those skilled in the art according to known techniques.

[0161] The fusion antigen polypeptides shown in SEQ ID NOs: 48-51 (referred to as fusion antigen polypeptides 1, 2, 3, and 4) were synthesized by Merrifield solid-phase synthesis method.

[0162] 1. Polypeptide synthesis

[0163] The synthesis order of Merrifield solid-phase synthesis is from C-termi...

Embodiment 2

[0182] Example 2. Preparation of Antigen Polypeptide Vaccine

[0183] 1. Water phase preparation

[0184] The fusion antigen polypeptides 1, 2, 3, and 4 obtained in Example 1 were diluted to 50 μg / ml with sterilized water for injection, and filtered through a filter with a pore size of 0.2 μm.

[0185] 2. Preparation of oil phase

[0186] The oil phase adjuvant MontanideISA50V2 was sterilized at 120°C for 30 minutes before use.

[0187] 3. Emulsification

[0188] First add the oil phase into the emulsification tank, then stir at 80-100r / min, and at the same time slowly add the water phase, the ratio of the oil phase / water phase is 1 / 1, stir for 2 minutes after adding, and then stir at 8500r / min After 6 minutes, it was emulsified to form a water-in-oil emulsion to obtain the foot-and-mouth disease virus vaccine of the present invention, which was recorded as vaccine 1, 2, 3, and 4.

Embodiment 3

[0189] Example 3. Detection of antibody levels after different doses of antigen immunization

[0190] 1. Test material

[0191] Test samples: vaccines 1, 2, 3, and 4 prepared in Example 2.

[0192] Experimental animals: healthy pigs with negative foot-and-mouth disease, weighing about 40kg / head, about 4 months old.

[0193] 2. Test method

[0194] Take 25 μg, 8.33 μg and 2.78 μg of vaccines 1, 2, 3, and 4 formulated with fusion antigen polypeptides, and inoculate 20 pigs once in each dose group by intramuscular injection behind the root of the ear, respectively, on the 7th, 14th, 21st, and 28th day Blood was collected, serum was separated, and the antibody level after antigen immunization was detected by liquid phase blocking enzyme-linked immunosorbent assay (LPB-ELISA) method for FMD A antibody. The basic principle of the liquid-phase blocking enzyme-linked immunosorbent assay is as follows: coat the ELISA plate with rabbit anti-foot-and-mouth disease virus antibody, tr...

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Abstract

The present invention discloses an A-type antigen polypeptide, a fusion antigen polypeptide and a vaccine of a foot and mouth disease virus, in particular relating to an A-type antigen polypeptide and a fusion antigen polypeptide of the foot and mouth disease virus, and a foot and mouth disease virus vaccine containing the antigen polypeptide and / or a fusion antigen polypeptide. The present invention also provides preparation methods of the antigenic polypeptide, the fusion antigen polypeptide and the vaccine. The present invention further provides applications of the antigen polypeptide, the fusion antigen polypeptide and the vaccine in preventing and controlling of the foot and mouth disease virus infection. The A-type antigen polypeptide, the fusion antigen polypeptide and the vaccine of the foot and mouth disease virus disclosed by the present invention have a broad spectrum immunogenicity, and may produce good immunogenicity on different foot and mouth disease viruses and variants thereof.

Description

technical field [0001] The invention relates to a foot-and-mouth disease virus antigen polypeptide, a fusion antigen polypeptide and a vaccine, specifically a foot-and-mouth disease virus type A antigen polypeptide, a fusion antigen polypeptide, and a foot-and-mouth disease virus type A vaccine containing the above-mentioned antigen polypeptide and / or fusion antigen polypeptide. The present invention also relates to the preparation and application of the above-mentioned antigen polypeptide, fusion antigen polypeptide and vaccine. Background technique [0002] Foot-and-mouth disease is an acute, febrile, highly contagious zoonotic disease caused by foot-and-mouth disease virus. Foot-and-mouth disease mainly affects cloven-hoofed animals, and its clinical diagnosis is characterized by blisters on the oral mucosa, hooves and breast skin. Adult animals infected with foot-and-mouth disease generally have a low mortality rate, but the morbidity rate can reach 100%. A large number...

Claims

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Application Information

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IPC IPC(8): C07K14/09C12N15/42C07K19/00A61K39/135A61P31/14G01N33/569
CPCA61K39/12A61K2039/545A61K2039/552A61K2039/55566A61K2039/70C07K14/005C07K2319/00C12N2770/32122C12N2770/32134G01N33/56983
Inventor 陈智英郑海学刘湘涛张震王江辉才学鹏聂东升殷宏
Owner SHANGHAI SHEN LIAN BIOMEDICAL CORP
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