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Vaccine composition and its preparation method and use

A vaccine composition and protein antigen technology, applied in the field of veterinary biological products, can solve the problems of large-scale production difficulty, low efficiency of all toxins, and high production costs

Active Publication Date: 2016-07-27
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the secretion of natural PMT is very limited, less than 0.6% of bacterial protein, and the purification process is complicated, and the efficiency of artificially expressing the whole toxin is also low. Therefore, the production cost of preparing toxoid seedlings by purifying natural PMT is high, and large-scale production High difficulty

Method used

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  • Vaccine composition and its preparation method and use
  • Vaccine composition and its preparation method and use
  • Vaccine composition and its preparation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The construction of embodiment 1 bacterial classification

[0054] 1.1 Construction of engineering strains expressing N-terminal and C-terminal proteins of Bordetella bronchiseptica PRN adhesin protein

[0055] 1.1.1 Primer design

[0056] Referring to the prn gene sequence (AJ245927) published on GenBank, two pairs of primers were designed using Primer5.0 software.

[0057] Table 1 prn gene 5' end and 3' end amplification primer sequence

[0058] Primer

Primer sequence

restriction endonuclease

prn F1

5'-CGC GGATCC AACATGTCTCTGTCACGCATTGTC-3'

BamHI

prn R1

5'-CCG CTCGAG GATATCGACCTTGCCGTCCTT-3'

wxya

prn F2

5'-CCG GAATTC GGTACCTACCGCTATCGATTG-3'

EcoR I

prn R2

5'-CCC AAGCTT CCAGCTGTACCGGTAGCC-3'

Hind III

[0059] Note: The underlined part is the enzyme cutting site

[0060] 1.1.2 Construction of rPRN-N and rPRN-C expression engineering strains

[0061] According to the kit i...

Embodiment 2

[0080] The preparation of embodiment 2 antigen

[0081] 2.1 Preparation of N-terminal and C-terminal proteins of Bordetella bronchiseptica PRN adhesin protein

[0082] The engineered strains BL21-PRNN and BL21-PRNC were respectively streaked on LB plates (30 μg / ml) containing kanamycin and cultured in a 37° C. incubator for 16 hours. Single colonies of BL21-PRNN and BL21-PRNC were picked respectively, inoculated in 5 ml of LB liquid medium containing kanamycin (final concentration 30 μg / ml), and cultured on a shaker (200 rpm) at 37°C for 16 hours to 1% of the ratio was inoculated in LB liquid medium containing 30 μg / ml kanamycin at 37°C on a shaker (200 rpm) and cultivated to OD 600 When it reaches 0.8-1.0, add an appropriate amount of isopropylthio-β-D-galactoside (IPTG) and continue culturing for 3-4 hours. At the end of induction, heat in a water bath, inactivate at 60°C for 60 minutes, and shake once every 5 minutes during this period. After inactivation, samples were t...

Embodiment 3

[0089] The preparation of embodiment 3 vaccines

[0090] Take the prepared rPRN-N, rPRN-C, rPMT-N and rPMT-C protein antigens, add an appropriate amount of MontanideGEL01 adjuvant, stir with a magnetic stirrer, and finally add thimerosal so that the final concentration of thimerosal is 1 / 10,000 1. Mixing, testing, and subpackaging, it is the porcine atrophic rhinitis subunit vaccine, wherein the content of each antigen in the vaccine is 12.5-50 μg / ml. The specific composition of each embodiment is as shown in Table 3:

[0091] Table 3 Specific implementation cases of vaccine ratio

[0092]

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Abstract

The invention provides a vaccine composition. The vaccine composition comprises the immune dose of Bordetella bronchiseptica pertactin (PRN) protein end N and end C, toxigenic Pasteurellamultocida Pasteurellamultocidatoxin (PMT) toxin protein end N and end C and pharmaceutically acceptable carriers. The vaccine composition has good immunogenicity and can effectively prevent and / or treat atrophic rhinitis of a pig. The invention provides a preparation method of the vaccine composition. Through gene engineering means, a lot of protein antigens are expressed. The vaccine composition has good safety and is conducive to large scale production.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, and in particular relates to a porcine atrophic rhinitis vaccine composition, a preparation method and application of the vaccine. Background technique [0002] Atrophic rhinitis (atrophicrhinitis, AR) is an important infectious disease of the respiratory system in pigs. It can cause symptoms such as chronic rhinitis, sneezing, tearing, facial degeneration and turbinate atrophy in pigs. The snout is deformed and affects the bone development of the whole body, which leads to growth retardation of pigs, makes pigs susceptible to secondary complex pneumonia, and causes huge losses to the pig industry. [0003] Porcine atrophic rhinitis can be divided into progressive atrophic rhinitis (PAR) and nonprogressive atrophicrhinitis (NPAR). [0004] Non-progressive porcine atrophic rhinitis is simply caused by Bordetella bronchiseptica (Bb), which usually only causes mild turbinate atrophy, a...

Claims

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Application Information

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IPC IPC(8): A61K39/385A61P11/02
Inventor 张许科孙进忠谷世江田克恭
Owner PU LIKE BIO ENG
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