Polypeptide used for preparing O type peptide vaccine of cattle foot-and-mouth disease and preparation methods and applications thereof

A bovine foot-and-mouth disease and peptide vaccine technology, applied in the field of medicine, can solve the problems of not being able to effectively protect animals, affecting the use of new vaccines, poor effect, etc., and achieve the effect of good immune efficacy, good application prospects, and no biological safety.

Active Publication Date: 2013-07-03
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 engi

Method used

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  • Polypeptide used for preparing O type peptide vaccine of cattle foot-and-mouth disease and preparation methods and applications thereof
  • Polypeptide used for preparing O type peptide vaccine of cattle foot-and-mouth disease and preparation methods and applications thereof
  • Polypeptide used for preparing O type peptide vaccine of cattle foot-and-mouth disease and preparation methods and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Solid-phase synthesis of bovine foot-and-mouth disease O-type synthetic peptide antigen

[0063] 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. Rink Amide MBHA resin from Sigma. The production process usually includes solid-phase synthesis of polypeptide antigens, cleavage of polypeptides, purification of antigens and sterilized storage.

[0064] 1.1 Solid-phase synthesis of synthetic peptide antigens

[0065] 1.1.1 Preparation of synthetic raw materials

[0066] The sequences of the synthetic polypeptide antigens are respectively shown in SEQ ID NO.5, SEQ ID NO.6 or SEQ ID NO.7.

[0067] According to the sequence of the antigen and the synthesis scale of 1 mmol, prepare the appro...

Embodiment 2

[0093] Example 2 Preparation of Synthetic Peptide Vaccine

[0094] 2.1 Preparation of antigen aqueous phase

[0095] Weigh the synthetic peptide antigens prepared according to Example 1 whose sequences are respectively shown in SEQ ID NO.5, SEQ ID NO.6 or SEQ ID NO.7, and then dilute the concentration of the synthetic peptide antigen to 50 μg with sterile water for injection / ml. The resulting antigen solution was filtered through a filter with a pore size of 0.2 μm and sterilized.

[0096] 2.2 Preparation of oil phase adjuvant

[0097] Sterilize the oil phase adjuvant at 50V at 121°C for 30 minutes, and set aside.

[0098] 2.3 Emulsion of synthetic peptide vaccine

[0099] 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 at a slow speed of 90-...

Embodiment 3

[0100] Example 3 Efficacy Test of Bovine Foot-and-Mouth Disease Type O Synthetic Peptide Vaccine

[0101] 1. Materials and methods

[0102] 1.1 Synthetic peptide vaccine

[0103] According to Example 1, prepare the polypeptide antigen whose sequence is shown in SEQ ID NO.5, SEQ ID NO.6 or SEQ ID NO.7, and then prepare the corresponding batch numbers according to Example 2: ZM01, ZM02 and ZM03 O-type Foot-and-mouth disease synthetic peptide vaccine.

[0104] 1.2 Experimental animals

[0105] 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).

[0106] 1.3 Virus Seed OS / 99

[0107] 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.

[0108] 1.4 Test method

[0109] Five cattle were immunized with each of the three groups of peptide vaccines tested, and a conventional inactivate...

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Abstract

The invention provides polypeptide used for preparing O type peptide vaccine of cattle foot-and-mouth disease. The polypeptide comprises an amino acid sequence presented by SEQ ID NO.1 and the peptide vaccine containing the polypeptide. The O type synthetic peptide vaccine of cattle foot-and-mouth disease provided by the invention has excellent immune efficacy, and cannot cause the problems of fever, swelling and the like existing in traditional vaccines, so that the vaccine can effectively respond to antigenic variation of current foot-and-mouth disease virus, prevents biosecurity problems, is easy to synthesize in a large scale, and has an excellent application prospect. The invention further provides preparation methods and pharmaceutical applications 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 O-type peptide vaccine, a vaccine containing the polypeptide 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 huge e...

Claims

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Application Information

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IPC IPC(8): C07K14/09C07K1/34C07K1/06C07K1/04A61K39/135A61P31/14
CPCY02P20/55
Inventor 巴利民齐鹏肖进栗利芳宋芳董春娜赵洪涛郑应华郑明举张爱民马爱荣
Owner 中国牧工商集团有限公司
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