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Preparation and application of fusion protein and vaccine composition thereof

A vaccine composition and fusion protein technology, applied in the field of veterinary biological products, can solve problems such as sudden onset, high morbidity and mortality, and economic loss

Active Publication Date: 2015-01-07
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease has a sudden onset, rapid spread, wide epidemic range, and long epidemic time. It is ineffective to use various antibiotics for prevention and treatment. Porcine Epidemic Diarrhea Virus Variant (Chen JF, Liu XZ, Shi D, et al. Complete Genome Sequence of a Porcine Epidemic Diarrhea Virus Variant. Journal of Virology, 2012, 86:3408)
[0004] At present, the prevention and control of the disease mainly adopts traditional techniques to prepare inactivated vaccines for immunization, but the whole PEDV virus is difficult to isolate and culture.

Method used

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  • Preparation and application of fusion protein and vaccine composition thereof
  • Preparation and application of fusion protein and vaccine composition thereof
  • Preparation and application of fusion protein and vaccine composition thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Preparation, identification and content determination of fusion protein 1

[0074] According to the COE neutralization table of the S protein fragment in the porcine epidemic diarrhea virus (accession number: JN599150.1, AB857233.1, AB857234.1) reported in NCBI (http: / / www.ncbi.nlm.nih.gov) The gene sequence of the position (see SEQ ID No. 1, SEQ ID No. 3 (artificial synthesis), SEQ ID No. 5, SEQ ID No. 7), the gene sequence of the neutralizing epitope of the combination of SS2-SS6-2C10 (SEQ ID No. 9), and the gene sequence of the Fc protein in porcine immunoglobulin IgG (accession number: NM_213828.1) (see SEQ ID No. 11) to prepare the fusion protein by genetic engineering.

[0075] 1.1 Construction and identification of antigenic protein of porcine epidemic diarrhea virus

[0076] When constructing the COE neutralizing epitope in the S protein fragment of porcine epidemic diarrhea virus, specific restriction sites such as EcoRI and HindIII were introduced at the 5...

Embodiment 2

[0091] Example 2 Preparation of porcine epidemic diarrhea virus vaccine composition

[0092] Use pH 7.4PBS to dilute the fusion protein prepared in Example 1, and add aluminum gel adjuvant to mix well so that the content of the recombinant fusion protein contained in the vaccine composition is shown in Table 1, while ensuring the aluminum gel adjuvant The volume ratio to the vaccine composition is 1:5. The prepared vaccine composition is used as an immunogen and stored at 4°C for later use.

[0093] Table 1 Components contained in porcine epidemic diarrhea vaccine composition

[0094]

Embodiment 3

[0095] Example 3 Evaluation of immune efficacy of porcine epidemic diarrhea vaccine composition

[0096] 3.1 Active immunity test

[0097] A total of 64 3-day-old PEDV antibody and antigen-negative piglets were selected and randomly divided into 8 groups (see Table 2 for details), 8 pigs per group. Groups 1-1, 1-2, 1-3, 1-4, 1-5, and 1-6 were injected intramuscularly with the vaccine prepared in Example 2 1-5, 2ml / head; No. 1-7, 1- The 8 groups were all blank control groups immunized with the same dose of pH7.4 PBS solution. On the 14th day after immunization, sera from the immunized group and the control group were collected, and the serum neutralizing antibody titer of each group was detected. The SPSS 15.0 software was used for statistical analysis of the average antibody titer of each piglet. At the same time, in the immune group, except for the 1-1, 1-5 and blank control groups, the pigs in groups 1-7 were given 1ml of Shanghai virus strain virus liquid (viral liquid TCID 5...

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Abstract

The invention provides a fusion protein, a porcine epidemic diarrhea vaccine composition containing the fusion protein and application thereof. The fusion protein contains porcine epidemic diarrhea virus antigenic protein and immunoglobulin Fc segment, wherein the porcine epidemic diarrhea virus antigenic protein contains a protein formed by series combination of porcine epidemic diarrhea virus S protein segments. The invention also provides a porcine epidemic virus vaccine composition which contains the fusion protein and a carrier. The invention also provides a preparation method of the vaccine composition and application of the vaccine composition in preparing drugs for preventing and / or treating diseases initiated by porcine epidemic diarrhea virus. The vaccine composition prepared from the fusion protein avoids the technical problem that the porcine epidemic diarrhea virus totivirus can not be easily separated and cultured in the traditional vaccine inactivation process. The fusion protein can utilize the gene engineering technique to perform abundant recombinant expressions, has the advantage of short time consumption, and is convenient for large-scale production.

Description

Technical field [0001] The invention belongs to the technical field of veterinary biological products, and specifically relates to the preparation and application of a fusion protein of porcine epidemic diarrhea virus and a vaccine composition containing the fusion protein. Background technique [0002] Porcine epidemic diarrhoea (PED) is an important acute, contact, viral intestinal infectious disease caused by porcine epidemic diarrhea virus (PEDV). It is watery diarrhea, It is characterized by vomiting, dehydration and decreased appetite. The disease can occur in pigs of any age. The younger the age, the more severe the symptoms, and the higher the mortality rate. In particular, newborn piglets within 7 days of age will experience severe dehydration and die 3-4 days after diarrhea occurs. The mortality rate is as high as 50. %, the highest mortality rate is 100%. The disease first occurred in the United Kingdom and Belgium in the 1970s, and was isolated to PEDV in Shanghai, ...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K39/215A61K47/48A61P31/14
Inventor 张许科孙进忠王延辉田克恭
Owner PU LIKE BIO ENG
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