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Polypeptide for preparing cattle foot-and-mouth disease ASIAI type peptide vaccine and preparing method and use thereof

A technology of bovine foot-and-mouth disease and peptide vaccines, applied in the field of medicine, can solve problems such as ineffective protection of animals, impact on the use of new vaccines, poor efficacy, etc.

Active Publication Date: 2013-07-31
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 cattle foot-and-mouth disease ASIAI type peptide vaccine and preparing method and use thereof
  • Polypeptide for preparing cattle foot-and-mouth disease ASIAI type peptide vaccine and preparing method and use thereof
  • Polypeptide for preparing cattle foot-and-mouth disease ASIAI type peptide vaccine and preparing method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Solid-phase synthesis of ASIA type I synthetic peptide antigen of bovine foot and mouth disease

[0075] The synthetic peptide antigen of the present invention can be prepared using ABI 433 type automatic peptide synthesizer by Merrifield solid-phase synthesis method, in which 9-fluorenylmethyloxycarbonyl (Fmoc) modified amino acids are used, and the solid-phase carrier is purchased from the United States Rink Amide MBHA resin from Sigma. The production process usually includes solid-phase synthesis of peptide antigens, peptide lysis, antigen purification and sterilization preservation.

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

[0077] 1.1.1 Preparation of synthetic raw materials

[0078] The sequence of the synthetic polypeptide antigen is 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, respectively.

[0079] According to the sequence of the antigen and the synthesis scale of 1 mmol, prepare s...

Embodiment 2

[0105] Example 2 Preparation of synthetic peptide vaccine

[0106] 2.1 Preparation of antigen aqueous phase

[0107] Weigh the sequences prepared in accordance with Example 1 as 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, respectively The peptide antigen is synthesized, and then the concentration of the synthesized peptide antigen is diluted to 50 μg / ml with sterile water for injection. The obtained 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] The oil phase adjuvant 50V was sterilized at 121°C for 30 minutes, and then used.

[0110] 2.3 Emulsification of synthetic peptide vaccine

[0111] Clean the IKA emulsification equipment 3 times with 2000ml of sterilized distilled water, and then add the oil phase to the emulsification tank at a volume ratio of 1:1 between the oil phase adjuvant and the antigen aqueous phase at 20-28°C, and start t...

Embodiment 3

[0112] Example 3 Efficacy Test of Bovine Foot-and-Mouth Disease ASIA Type I Synthetic Peptide Vaccine

[0113] 1. Materials and methods

[0114] 1.1 Synthetic peptide vaccine

[0115] According to Example 1, the polypeptide antigens 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 were prepared according to The corresponding batch numbers prepared in Example 2 were: ZM62601, ZM62602, ZM62603, ZM62604, ZM62605, ZM62606 ASIA type I foot-and-mouth disease synthetic peptide vaccine.

[0116] 1.2 Experimental animals

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

[0118] 1.3 Virus species ASIAI JSL

[0119] Measure and adjust the toxic value with 3-4 day-old suckling mice, and store them at -25℃ for later use.

[0120] 1.4 Test method

[0121] Each of the 6 groups of test peptide vacci...

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PUM

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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 present invention belongs to the field of medical technology. Specifically, the present invention relates to a polypeptide used for preparing a bovine foot-and-mouth disease ASIA type I peptide vaccine, as well as vaccines containing these polypeptides and their preparation methods. Background technique [0002] Foot and mouth disease (foot and mouth disease, FMD for short) is an acute, highly contagious, and febrile infectious disease that occurs in cloven-hoofed animals and is widely distributed worldwide. Foot-and-mouth disease is highly contagious and spreads rapidly. After infecting pigs, cattle, sheep and other livestock, it will cause the death of young animals, and the production capacity of adult animals will decline sharply. Therefore, it seriously endangers the development of animal husbandry and the production and supply of meat and animal products. At present, the foot-and-mouth disease has greatly blocked and restricted the market circulat...

Claims

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Application Information

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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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