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C-shaped aftosa subunit vaccine and preparation method thereof

A subunit vaccine, foot-and-mouth disease virus technology, applied in the field of vaccines and their preparation, can solve problems such as inconvenience and safety problems, and achieve the effects of high immune protection and good immune effects

Active Publication Date: 2014-05-28
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The use of inactivated vaccines is still the main means of controlling FMD, but there are safety problems and inconveniences in production, transportation and use, so the development of new vaccines is an effective way to solve these problems

Method used

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  • C-shaped aftosa subunit vaccine and preparation method thereof
  • C-shaped aftosa subunit vaccine and preparation method thereof
  • C-shaped aftosa subunit vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 Amplification of type C foot-and-mouth disease antigen gene VP1 and VP3 and expression in Chinese hamster ovary cell expression system

[0039] 1. Ligation and transformation of target gene and plasmid

[0040] VP1 primer

[0041] Upstream primer: 5′-GC GAATTC ACCACGACCACTGGTGAATCT-3′ (underlined as EcoR I restriction site)

[0042] Downstream primer: 5′-GC CTCGAG CTGCTTCGCGGGCGCGATGAG-3' (underlined as Xho I restriction site);

[0043] and VP3 primers

[0044] Upstream Primer::5′-GC GAATTC GGAATCTTTCCCGTCGCGTGC-3′ (underlined as Xho I restriction site)

[0045] Downstream Primer::5′-GC CTCGAG TTGTTGTCTAGCGTCCACCG-3' (underlined as Xho I restriction site);

[0046] Amplify with type C foot-and-mouth disease virus as a template, including: genome template 100ng, dNTP (10mM) 5μl, 10×pfu buffer 5μl, pfu DNApolymerase 5U and VP1 primer or VP3 primer forward and reverse primer 50pmol each, supplemented with sterile ultra-thin Pure water to a total vol...

Embodiment 2

[0076] Embodiment two: the preparation of C type foot-and-mouth disease subunit genetic engineering vaccine;

[0077] In this implementation, the eukaryotic expression products of VP1 and VP3 obtained above are mixed with 206 oil adjuvant to make a vaccine:

[0078] (1) The eukaryotic expression products of VP1 and VP3 were respectively diluted to 50 μg / ml with sterile water, and the water phase.

[0079] (2) Mix the diluted VP1 and VP3 with a protein content of 1:1, then mix the resulting mixture of VP1 and VP3 with 206 oil adjuvant at a volume ratio of 1:1, first at a slow speed of 90-150r / m After rotating and stirring for 2 minutes, stir at a high speed of 10000r / m for 20 minutes, and let stand for 5 minutes to make a vaccine.

[0080] (3) Divide the obtained emulsion under aseptic conditions to obtain the type C foot-and-mouth disease genetic engineering subunit vaccine.

Embodiment 3

[0081] Embodiment three: the effect test of C type foot-and-mouth disease subunit genetic engineering vaccine (comprising the immune antibody determination of mouse, guinea pig, pig; The challenge test of guinea pig is measured)

[0082] Experiment 1 above-mentioned a kind of C type foot-and-mouth disease genetic engineering subunit vaccine immunity test, specifically as follows:

[0083] 1.1 Materials and methods

[0084] 1.1.1 The vaccine is the type C foot-and-mouth disease genetically engineered subunit vaccine prepared according to Example 2, named batch numbers: 20110305, 20110320, 20110401.

[0085] As a control group:

[0086] C-type VP1 foot-and-mouth disease genetic engineering subunit vaccine was prepared by this experiment, named batch number: 20110211; C-type VP3 foot-and-mouth disease genetic engineering subunit vaccine was prepared by this experiment, named batch number: 20110221; C-type foot-and-mouth disease inactivated vaccine was prepared by this experiment...

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Abstract

The invention discloses a C-shaped aftosa subunit genetic engineering vaccine and a preparation method thereof. The C-shaped aftosa subunit vaccine has the main component of a mixture formed by C-shaped aftosa structure protein VP1 and VP3 according to the protein content ratio being 1 / 1, wherein the C-shaped aftosa structure protein VP1 and VP3 is eukaryotic expression products of C-shaped aftosa virus gene VP1 and VP3, in particular to products of C-shaped aftosa antigenic gene VP1 and VP3 expressed in a Chinese hamster ovarian cell (CHO / dhfr-cell) expression system.

Description

technical field [0001] The invention relates to a vaccine and a preparation method thereof. The vaccine involved in the invention is a type C foot-and-mouth disease vaccine and a preparation method thereof, in particular a type C foot-and-mouth disease subunit genetic engineering vaccine and a preparation method thereof. Background technique [0002] Foot-and-Mouth Disease (FMD) is an acute, severe and highly contagious infectious disease that can infect a variety of cloven-hoofed animals including cattle, sheep, pigs and other major livestock. Virus (Foot-and-Mouth Disease Virus, FMDV). FMDV belongs to the family Picornaviridae and the genus Aphthovirus, including seven serotypes: A, C, O, AsiaI, SAT I, SAT II and SAT III, and there is no cross-protection among the serotypes. The virions of FMDV have a diameter of 20-25nm, an icosahedral structure, and are approximately circular; the genome is a positive-strand RNA with a full length of 8500 (nt), including an open reading...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/135C12N15/85A61P31/14C12R1/93
Inventor 刘永生张杰丁耀忠陈豪泰马丽娜周建华
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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