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Preparation and application of a fusion protein and vaccine composition thereof

A vaccine composition and fusion protein technology, applied in the field of veterinary biological products, can solve the problems of high morbidity and mortality, difficulty in isolating and cultivating PEDV whole virus, and economic loss

Active Publication Date: 2017-07-28
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 a fusion protein and vaccine composition thereof
  • Preparation and application of a fusion protein and vaccine composition thereof
  • Preparation and application of a fusion protein and vaccine composition thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0074] According to the COE neutralization table in the S protein fragment of porcine epidemic diarrhea virus (accession number: JN599150.1, AB857233.1, AB857234.1) reported in NCBI (http: / / www.ncbi.nlm.nih.gov) (respectively see SEQ ID No.1, SEQ ID No.3 (artificially synthesized), SEQ ID No.5, SEQ ID No.7), the gene sequence of the neutralizing epitope of SS2-SS6-2C10 combination ( 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 means of genetic engineering.

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

[0076] When constructing the COE neutralizing epitope in the S protein fragment of porcine epidemic diarrhea virus, specific enzyme cutting sites such as EcoRI and HindIII were introduced at the 5'-end and 3'-e...

Embodiment 2

[0091] The preparation of embodiment 2 porcine epidemic diarrhea virus vaccine composition

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

[0093] The components contained in the porcine epidemic diarrhea vaccine composition of table 1

[0094]

Embodiment 3

[0095] Example 3 Porcine epidemic diarrhea vaccine composition immune efficacy evaluation

[0096] 3.1 Active immunization test

[0097] Select 64 PEDV antibody and antigen-negative piglets at the age of 3 days, and randomly divide them into 8 groups (see Table 2 for details), 8 pigs / group. Groups 1-1, 1-2, 1-3, 1-4, 1-5, and 1-6 were injected intramuscularly with vaccines prepared in Example 2 respectively at 1-5, 2ml / head; groups 1-7, 1- The 8 groups were blank control group and were immunized with the same dose of PBS solution with pH 7.4. On the 14th day after immunization, the sera of pigs in the immunized group and the control group were collected, and the titers of neutralizing antibodies in the sera of each group were detected. The average antibody titer of each piglet was statistically analyzed using SPSS 15.0 software. At the same time, pigs in the immunization group except groups 1-1, 1-5 and groups 1-7 of the blank control group were orally administered with 1ml...

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Abstract

The invention provides a fusion protein, 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 fragment, and the porcine epidemic diarrhea virus antigenic protein contains a protein formed by series combination of porcine epidemic diarrhea virus S protein fragments. The invention also provides a porcine epidemic virus vaccine composition, which contains fusion protein and carrier. The invention also discloses the preparation method of the vaccine composition and its application in the preparation of medicines for preventing and / or treating diseases caused by porcine epidemic diarrhea virus. The vaccine composition prepared by the fusion protein avoids the technical difficulty of separating and cultivating the whole virus in the process of preparing the traditional inactivated vaccine with the whole virus of porcine epidemic diarrhea virus; It is time consuming and also facilitates mass production.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular 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 diarrhea (porcine epidemic diarrhea, PED) is an important acute, contact, viral intestinal infectious disease caused by porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV). Vomiting, dehydration, and decreased appetite are characteristic. 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 die of severe dehydration 3-4 days after diarrhea occurs, and 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. In 1973,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00A61K39/215A61K47/68A61P31/14
Inventor 张许科孙进忠王延辉田克恭
Owner PU LIKE BIO ENG
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