Porcine reproductive and respiratory syndrome virus-like particle vaccine and preparation method thereof

A technology for respiratory syndrome and pig reproduction, applied in biochemical equipment and methods, antiviral agents, viruses/phages, etc., can solve the problems of no potential danger, long preparation period and high cost

Inactive Publication Date: 2011-06-29
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, compared with live vaccines and inactivated vaccines, VLP vaccines have no unsafe factors; compared with subunit vaccines, VLPs are composed of multiple antigenic proteins and have strong immunogenicity; Compared with Virosome, VLP is neither prepared from live virus, has no potential danger, does not require a large number of reproductive viruses, and has no disadvantages such as low safety, high cost, and long preparation cycle; thus VLP vaccines are widely used. Today is hailed as a new milestone in the development of viral vaccines

Method used

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  • Porcine reproductive and respiratory syndrome virus-like particle vaccine and preparation method thereof
  • Porcine reproductive and respiratory syndrome virus-like particle vaccine and preparation method thereof
  • Porcine reproductive and respiratory syndrome virus-like particle vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0030] Experimental example 1: PRRS VLP vaccine verification experiment of the present invention

[0031] The PRRS VLP vaccine injection, nasal drops, and drinking water formulations prepared in Example 8 of the present invention are uniform in appearance, without peculiar smell and bacterial contamination; they have no pyrogen, abnormal toxicity and acute toxicity through guinea pig and rabbit tests; Intracerebral inoculation in rats and intramuscular injection in the neck of 3-day-old piglets showed no signs of disease; the protein content determined by the Lorry method was within the dose range; the diameter of VLP was observed by electron microscopy to be about 45-65nm, which was consistent with natural viruses; the DNA content was determined by ELISA All were below 50EU; the presence of antigenic protein components was detected by SDS-PAGE and WB; the neutralizing antibody titer in the serum of Balb / C mice once immunized was above 3200.

experiment example 2

[0032] Experimental Example 2: Effect experiment of the PRRS VLP vaccine of the present invention on the immune response in experimental pigs

[0033]The PRRS VLP vaccine prepared in Example 8 of the present invention was added or not added to the corresponding adjuvant prepared injection, nasal drip, and drinking water vaccine dosage forms, and PBS, aluminum salt, nano-aluminum, P3BSK4, rLT, were used as the control group, and were immunized according to their respective routes. 3 days old piglets, and its immunization dosage of back-up sows are immunized twice (0, 21 days) by embodiment 8 vaccine low dosage group; Collect the peripheral blood of each group piglets and sows 14 days after the last immunization, separate For serum and immune cells, spleen lymphocytes, lung and nasal lavage fluid were collected; the serum antibody titer was determined to be over 6400 by ELISA method, and the neutralizing antibody titer was determined to be over 3200 by MTT method, and was measure...

experiment example 3

[0034] Experimental Example 3: Experiment of the immune protection effect of the PRRS VLP vaccine of the present invention on experimental pigs

[0035] The PRRS VLP vaccine prepared in Example 8 of the present invention was prepared with or without adding corresponding adjuvants in the form of injection, nasal drops, and drinking water as the experimental group, and the virus was inactivated with PBS, aluminum salt, nano-aluminum, P3BSK4, rLT, and PRRSVCH-1a Be a control group, immunize piglets and gilts at the age of 3 days by their respective approaches, and their immunization doses are immunized twice (0, 21 days) by the vaccine low-dose group in Example 8, and each test group, The experimental pigs in the control group used 10 5.0 TCID 50 Clinically isolated PRRSV strains were challenged by intramuscular injection in the neck to observe the protective effect. The result shows that PRRS VLP vaccine of the present invention all can produce high-level protection effect, an...

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Abstract

A porcine reproductive and respiratory syndrome virus-like particle vaccine and a preparation method thereof relate to the field of biological medicines and aims at disclosing the porcine reproductive and respiratory syndrome virus-like particle vaccine (PRRS VLP vaccine) and the preparation method the vaccine. The PRRS VLP vaccine contains a VLP comprising three structure proteins of porcine reproductive and respiratory viruses M, N and GP5 and can excite favorable dual cell and humoral immune response. By adding or not adding an adjuvant into the formed VLP protein antigen, the pharmacodynamic test shows the prepared injection type, nasal drop type and water agent type vaccines are immunized with different animal groups so as to safely and effectively prevent the PRRSV infection and provide ideal vaccines for safely and effectively immunizing, preventing and controlling the PRRSV infection on different groups of sows, piglets and fat pigs.

Description

field of invention [0001] The invention belongs to the field of biopharmaceuticals, in particular to a porcine reproductive and respiratory syndrome virus-like particle vaccine (PRRS VLP vaccine) and a preparation method thereof. Background technique [0002] Porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome, PRRS) is a virus that causes abortion in sows and respiratory diseases in piglets. It was isolated in the Netherlands in 1991 and named Lelystad virus ( Lelystad Virus), a virus that can cause the same symptoms was also isolated in the United States in 1992, and then the virus was officially named porcine reproductive and respiratory syndrome virus, and the European strain represented by Lelystad Virus (LV) and Lelystad Virus (LV) were established. American strain represented by ATCC VR2332 strain. At the end of 1995, an epidemic with clinical manifestations of elevated body temperature and red skin, accompanied by high ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/295C12N15/85C12N7/00C12N7/02A61P31/14A61K39/12C12R1/93
Inventor 吕凤林曹政田菲菲杨力
Owner CHONGQING UNIV
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