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Preparation method for vaccine of porcine circovirus II

A technology for preparing porcine circovirus and vaccine, which is applied in the field of animal virus and porcine circovirus type II vaccine preparation, can solve the problems of affecting cell growth, high production cost, low single batch yield, etc., and achieves precise control of process parameters, The effect of product cost reduction and product quality stability

Inactive Publication Date: 2010-07-14
PU LIKE BIO ENG
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AI Technical Summary

Problems solved by technology

The disadvantages of this method are: stirring produces shear force, and bubbles are generated, affecting cell growth, and there are technical bottlenecks in large-scale production
(2) Rotating bottle technology: At present, this technology is basically used in industrial production. When producing cells and viruses, it is labor-intensive, occupies a large area, has large batch-to-batch differences, high production costs, and low single-batch yield, making it difficult to standardize the quality of production. control
[0005] In my country, there is no PCV2 vaccine that is really effective and approved by the Ministry of Agriculture. The main reason is that the antigen concentration is low, and the growth titer of PCV2 in cells is low, generally only 10 4.0 TCID 50 / ml or so

Method used

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  • Preparation method for vaccine of porcine circovirus II
  • Preparation method for vaccine of porcine circovirus II
  • Preparation method for vaccine of porcine circovirus II

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1 tidal type microcarrier suspension culture technology high-density cultivation of PK15 cells and porcine circovirus type II (PCV2)

[0033] The bioreactor used was TideCell-020 from CESCO, USA, and the carrier used was BioNocII polyester fiber from CESCO, USA.

[0034] (1) PK15 cells 4.0×10 9 The cell was inoculated in a 20L carrier tank and the carrier BioNocII polyester fiber 2.2×10 2 g, and DMEM growth medium containing 6% calf serum with a working volume of 500 L, cultivated by tidal microcarrier suspension culture technology until the total number of cells reaches 4.0×10 10 cell; when culturing cells, start the attachment program first, and switch to the culture program after 4 hours; the cell attachment program is: up: 2500mL / min hold 1min, Down: 2500mL / min hold 30s, set the maximum exchange volume of 18500mL; cell culture program : Up: 1900mL / min hold 1min, Down 1900mL / min hold 1min, set the maximum exchange volume of 18000mL;

[0035] (2) All the...

Embodiment 2

[0044] Preparation of inactivated porcine circovirus type 2 (PCV2) vaccine and evaluation of immune efficacy

[0045] 1. Materials and methods

[0046] 1.1 Vaccines

[0047] The porcine circovirus type II (PCV2) virus solution obtained in Example 1 was added with 206 adjuvant (French SEPPIC company product) at a ratio of 1:1, and fully mixed to get the final product (batch numbers are: 0803, 0804, 0805, 0806 and 0807 ). Finished product testing of the prepared vaccines to confirm compliance with various standards;

[0048] Character test: The seedling is a two-phase light red emulsion of water-in-oil-in-water, and the character test has reached the requirements of the "Quality Standard (Draft)". Take a clean straw, suck up a small amount of vaccine and drop it in cold water, all but the first drop will not spread. Stability test: no delamination and no demulsification when placed at 37°C for 21 days. Take 10ml of the vaccine and put it in a centrifuge tube, centrifuge at ...

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Abstract

The invention discloses mass production of inactivated vaccine of porcine circovirus II (PCV2) and a preparation method thereof. The preparation method comprises the following technical steps: (1) high-density culture of cells for vaccine preparation; (2) reproduction of venom for vaccine preparation; and (3) addition of adjuvant to prepare the inactivated vaccine. Compared with the prior art, the invention has the advantages that the virus yield is high, the virus titer is high, the production scale is large, the yield of a single batch is high, the production cost is relatively low, the product quality is high and stable, the operation is convenient, the operation space is small, the technological parameters are controlled accurately and the like. The inactivated vaccine has high safety, can induce pig bodies to generate immune protection and fully satisfies the national biological product standards.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, relates to animal viruses, and more particularly relates to a preparation method of porcine circovirus type II vaccine. Background technique [0002] Porcine circovirus type II (referred to as PCV2) infection caused by weaning piglet multisystemic wasting syndrome (PMWS), porcine respiratory disease syndrome (PRDC), porcine skin and nephrotic syndrome (PDNS) and other diseases (collectively referred to as porcine Circovirus diseases, PCVDs), are clinically characterized by progressive weight loss and signs such as tachypnea, dyspnea, and jaundice. From a pathological point of view, it manifests as lymphocytic or granulomatous infiltrates, lymphadenopathy and, less commonly, hepatitis and lymphocytic or granulomatous nephritis. (Clark E.G. (1997) Proc. Am. Assoc. Swine Prac.499-501; La Semaine Veterinaire No.26, supplement to La Semaine Veterinaire 1996 (834); La Semaine Ve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/20C12N7/00
CPCG01N33/56983G01N2333/01
Inventor 张许科孙进忠乔荣岑陶家权彭伍平张海洋习向锋李三
Owner PU LIKE BIO ENG
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