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Polypeptide fragment composition and application in preparation of porcine reproductive and respiratory syndrome vaccine thereof

A technology of polypeptide fragments and compositions, applied in the field of porcine reproductive and respiratory syndrome vaccine preparation, can solve problems such as difficult to control and eradicate PRRSV, strong virulence, not suitable for piglet immunization, etc., achieve good application prospects, and respond to antigens Variants, effects that are easy to synthesize on a large scale

Inactive Publication Date: 2012-01-18
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, while the attenuated vaccine provides immune protection, it will also continue to spread the vaccine virus, making the virus circulate in the pig farm, recombination and mutation may occur, and there are unsafe problems such as virulence reversion, and sometimes even PRRS break out
The inactivated vaccine has the disadvantages of large immunization dose, many immunization times, and long immunization cycle, and cannot provide immune protection to non-vaccine virus infection. It is not suitable for piglet immunization. important reason for being
Therefore, it is difficult to control and eradicate the existence and prevalence of PRRSV in pig farms through attenuated vaccines or inactivated vaccines

Method used

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  • Polypeptide fragment composition and application in preparation of porcine reproductive and respiratory syndrome vaccine thereof
  • Polypeptide fragment composition and application in preparation of porcine reproductive and respiratory syndrome vaccine thereof
  • Polypeptide fragment composition and application in preparation of porcine reproductive and respiratory syndrome vaccine thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1, the synthesis of polypeptide fragment and APP-N protein

[0035] 1. Synthesis of polypeptide fragments

[0036] Design six polypeptide fragments (all T cell epitopes), the sequence is as follows (from nitrogen terminal to carbon terminal):

[0037] Polypeptide I (sequence 1 of the sequence listing): TMPPGFELYGGGGSGGGGSFLLVGFKCF;

[0038] Polypeptide II (sequence 2 of the sequence listing): CLLPSLLAIGGGGSGGGGSLAALICFVIRLAKNC;

[0039] Polypeptide III (sequence 3 of the sequence listing): KGRLYRWRSPVIVEKGGGGSGGGGSCNDSTAPQKVLLAFS;

[0040] Polypeptide IV (sequence 4 of the sequence listing): ALKVSRGRLLGLLHLGGGGSGGGGSFGYMTFVHFESTNRV;

[0041] Polypeptide V (sequence 5 of the sequence listing): KFITSRCRLCLLGRKGGGGSGGGGSNPEKPHFPL;

[0042]Polypeptide VI (sequence 6 of the sequence listing): VRHHFPSEGGGGSGGGGSFSLPTQHTVRLIRATAS;

[0043] The above six polypeptides were chemically synthesized respectively.

[0044] 2. Synthesis of APP-N protein

[0045] APP-...

Embodiment 2

[0057] Embodiment 2, the preparation of vaccine

[0058] 1. Preparation of Vaccine A

[0059] 1. Dilute the polypeptide I, polypeptide II, polypeptide III, polypeptide IV, polypeptide V and polypeptide VI prepared in Example 1 to 1.2 mg / ml with water for injection (sterile water), and then mix them in equal volumes to make an aqueous phase Antigen (the concentration of the 6 polypeptide fragments in the aqueous phase antigen is 0.2mg / ml).

[0060] 2. Dilute the APP-N protein (adjuvant) prepared in Example 1 with water for injection (sterile water) to a protein concentration of 0.2 mg / ml, which is the aqueous phase of the protein adjuvant.

[0061] 3. Under the condition of 6°C (4-8°C is acceptable), mix the water-phase antigen in step 1 and the protein adjuvant in step 2 in equal volumes, and mix and combine at a slow speed of 50 rpm on a mixer After 2 hours, the aqueous phase adjuvant-antigen immunogen (the concentration of APP-N protein is 100μg / ml, the concentration of po...

Embodiment 3

[0071] Embodiment 3, efficacy experiment of vaccine

[0072] 1. Experimental animals

[0073] 50 6-week-old healthy pigs (Landrace pigs, purchased from Changping Animal Experiment Base, Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences) were double-negative for PRRSV antigen and antibody.

[0074] Second, the application of vaccines to immunize animals

[0075] The experimental animals were randomly divided into 5 groups, 10 in each group, and the following immunizations were carried out respectively:

[0076] Group 1 (experimental group A): the vaccine A prepared in Example 2 was used for immunization;

[0077] Group 2 (experimental group B): the vaccine B prepared in Example 2 was used for immunization;

[0078] The 3rd group (negative control group): adopt the reference substance A prepared in embodiment 2 to carry out immunization;

[0079] The 4th group (blank control group): adopt the reference substance B prepare...

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Abstract

The invention discloses a polypeptide fragment composition and an application in preparation of a porcine reproductive and respiratory syndrome vaccine thereof. The polypeptide fragment composition provided by the invention is a polypeptide shown in sequence 1 of the sequence table, a polypeptide shown in sequence 2 of the sequence table, a polypeptide shown in sequence 3 of the sequence table, apolypeptide shown in sequence 4 of the sequence table, a polypeptide shown in sequence 5 of the sequence table, and a polypeptide shown in sequence 6 of the sequence table. The polypeptide fragment composition provided by the invention can be used as an active component of a porcine reproductive and respiratory syndrome vaccine, and has great application value for the prevention and treatment of porcine reproductive and respiratory syndrome. The vaccine provided by the invention can effectively cope with antigenic variation of porcine reproductive and respiratory syndrome virus, has no biological safety problem, is suitable for large-scale synthesis, and has good application prospects.

Description

technical field [0001] The invention relates to a polypeptide fragment composition and its application in preparing porcine reproductive and respiratory syndrome vaccine. Background technique [0002] Porcine reproductive and respiratory syndrome (porcine reproductive and respiratory syndrome, PRRS), also known as porcine blue ear disease, was first discovered in the United States in 1987, and then spread rapidly to Canada. In Europe, the first outbreak occurred in Germany in 1990, followed by Norway, Belgium, Spain, the United Kingdom, France and Denmark. The disease has now spread to various parts of the world, including South America, North America, Europe and Asia. Recently, a highly virulent mutant strain has appeared in my country, which is a highly infectious disease of pigs caused by PRRS virus (PRRSV), which can cause reproductive disorders in sows, respiratory symptoms in pigs of all ages, and death of piglets. It can lead to immunosuppression and induce secondary...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K39/00A61K39/39A61P31/14
Inventor 刘文军陈才伟李晶杨利敏
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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