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Polypeptide for preparing bovine foot-and-mouth disease asia type I peptide vaccine and its preparation method and application

A technology of bovine foot-and-mouth disease and peptide vaccines, applied in the field of medicine, can solve the problems of ineffective protection of animals, poor effect, and influence on the use of new vaccines, and achieve good immune efficacy

Active Publication Date: 2015-12-23
中国牧工商集团有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In terms of research on new vaccines for foot-and-mouth disease, there have been reports on genetically engineered subunit vaccines, foot-and-mouth disease virus vector vaccines, and genetically engineered vaccines for foot-and-mouth disease. use of vaccines
In addition, these vaccines are often less effective against currently circulating strains that have mutated and cannot effectively protect animals

Method used

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  • Polypeptide for preparing bovine foot-and-mouth disease asia type I peptide vaccine and its preparation method and application
  • Polypeptide for preparing bovine foot-and-mouth disease asia type I peptide vaccine and its preparation method and application
  • Polypeptide for preparing bovine foot-and-mouth disease asia type I peptide vaccine and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] The solid-phase synthesis of embodiment 1 bovine foot-and-mouth disease ASIAI type synthetic peptide antigen

[0075] The synthetic peptide antigen of the present invention can be prepared by the Merrifield solid-phase synthesis method using the 433 type automatic polypeptide synthesizer of ABI Company, wherein amino acids modified by 9-fluorenylmethoxycarbonyl (Fmoc) are used, and the solid phase carrier is purchased from the United States. RinkAmideMBHA resin from Sigma. The production process usually includes solid-phase synthesis of polypeptide antigens, cleavage of polypeptides, purification of antigens and sterilized storage.

[0076] 1.1 Solid-phase synthesis of synthetic peptide antigens

[0077] 1.1.1 Preparation of synthetic raw materials

[0078] The sequences of the synthetic polypeptide antigens are respectively shown in SEQ ID NO.8, SEQ ID NO.9, SEQ ID NO.10, SEQ ID NO.11, SEQ ID NO.12 or SEQ ID NO.13.

[0079] According to the sequence of the antigen ...

Embodiment 2

[0105] The preparation of embodiment 2 synthetic peptide vaccines

[0106] 2.1 Preparation of antigen aqueous phase

[0107] Weigh the synthetic peptide antigens prepared according to Example 1 with sequences shown in SEQIDNO.8, SEQIDNO.9, SEQIDNO.10, SEQIDNO.11, SEQIDNO.12 or SEQIDNO.13, and then use sterile water for injection to synthesize The peptide antigen concentration was diluted to 50 μg / ml. The resulting antigen solution was filtered through a filter with a pore size of 0.2 μm and sterilized.

[0108] 2.2 Preparation of oil phase adjuvant

[0109] Sterilize the oil phase adjuvant at 50V at 121°C for 30 minutes, and set aside.

[0110] 2.3 Emulsion of synthetic peptide vaccine

[0111] Clean the IKA emulsification equipment with 2000ml of sterilized distilled water for 3 times, then put the oil phase into the emulsification tank at 20-28°C according to the volume ratio of oil phase adjuvant and antigen water phase at 1:1, and start the motor to After stirring a...

Embodiment 3

[0112] Embodiment 3 bovine foot-and-mouth disease ASIAI type synthetic peptide vaccine effectiveness test

[0113] 1. Materials and methods

[0114] 1.1 Synthetic peptide vaccine

[0115] According to Example 1, the polypeptide antigen whose sequence is shown in SEQIDNO.8, SEQIDNO.9, SEQIDNO.10, SEQIDNO.11, SEQIDNO.12 or SEQIDNO.13 is prepared, and then the corresponding batch numbers prepared according to Example 2 are: ZM62601, ASIAI type foot-and-mouth disease synthetic peptide vaccines of ZM62602, ZM62603, ZM62604, ZM62605, ZM62606.

[0116] 1.2 Test animals

[0117] 37 6-month-old healthy cattle (purchased from cattle farms in Lanzhou) were selected with negative foot-and-mouth disease antibody (nuclear mouse neutralizing antibody titer ≤ 1:4).

[0118] 1.3 Virus species ASIAIJSL

[0119] Use 3-4 day-old suckling mice to measure and adjust the toxicity, and store them in a freezer at -25°C for later use.

[0120] 1.4 Test method

[0121] Five cattle were immunized...

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Abstract

The invention provides polypeptide for preparing a cattle foot-and-mouth disease ASIAI type peptide vaccine. The polypeptide comprises an amino acid sequence shown as SEQIDNO.1 and the peptide vaccine comprising the polypeptide. The cattle foot-and-mouth disease ASIAI type combined peptide vaccine is good in immune potency and does not cause the problems of a fever, swelling and the like caused by a tradition vaccine. Thus, the polypeptide for preparing the cattle foot-and-mouth disease ASIAI type peptide vaccine can effectively solve the problems of antigen and variation and nonexistence of bio-security of an existing foot-and-mouth disease virus, can be easily synthesized massively and has a good application prospect. The invention further provides a preparing method and use of the polypeptide and the peptide vaccine.

Description

technical field [0001] The invention belongs to the technical field of medicine, in particular, the invention relates to a polypeptide used for preparing a bovine foot-and-mouth disease ASIAI type peptide vaccine, a vaccine containing these polypeptides and a preparation method thereof. Background technique [0002] Foot and mouth disease (FMD) is an acute, highly contagious, febrile infectious disease of cloven-hoofed animals, which is widely distributed in the world. Foot-and-mouth disease is highly contagious and spreads rapidly. After infecting pigs, cattle, sheep and other livestock, it will lead to the death of young animals and a sharp decline in the production capacity of adult animals. Therefore, it seriously endangers the development of animal husbandry and the production and supply of meat and animal products. At present, foot-and-mouth disease has greatly blocked and restricted the market circulation and international trade of animals and animal products, causing...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/09C07K7/08C07K1/06C07K1/04A61K39/135A61P31/14
CPCY02P20/55
Inventor 肖进齐鹏巴利民宋芳郑明举张欣张艳宾张君郑应华张爱民马爱荣
Owner 中国牧工商集团有限公司
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