Method for industrially producing swine parvovirus vaccine by utilizing bioreactor

A bioreactor and parvovirus technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of not being suitable for large-scale production of vaccines, high labor intensity in operation, and large differences between vaccine batches. Achieve the effects of saving manpower, high degree of automation control, and less production land

Active Publication Date: 2011-05-04
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, each spinner bottle is an independent cell culture unit, and the quality of cells, virus yield and titer in each bottle are different, resulting in large differences between vaccine batches, and the operation is

Method used

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  • Method for industrially producing swine parvovirus vaccine by utilizing bioreactor
  • Method for industrially producing swine parvovirus vaccine by utilizing bioreactor
  • Method for industrially producing swine parvovirus vaccine by utilizing bioreactor

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Bioreactor: 14L and 40L bioreactors from NBS Company of the United States;

[0021] Microcarrier: Cytodex-1 (General Electric Healthcare Life Sciences Division);

[0022] Porcine parvovirus: HW-1 strain;

[0023] Cell growth medium: DMEM containing 8% calf serum (Beijing Qingda Tianyi Biotechnology Co., Ltd.);

[0024] Virus maintenance solution: DMEM containing 1% calf serum by volume (Beijing Qingda Tianyi Biotechnology Co., Ltd.);

[0025] Cell culture: In 14L bioreactors, add Cytodex1 at a concentration of 10g / L, after hydration, wash with pH 7.2 phosphate buffer saline PBS twice, sterilize, inoculate IBRS-2 cells, and culture ; The parameters of the culture method are: pH7.2, temperature 37°C, DO50%, stirring speed 30-100rpm; take samples regularly every day to observe the growth of the cells, count the cells, and measure the consumption of glucose. When the density of the cells reaches 1.5×10 6 / ml, start perfusion, and the perfusion speed depends on the densit...

Embodiment 2

[0030] Bioreactor: 14L and 40L bioreactors from NBS Company of the United States;

[0031] Microcarrier: Cytodex-1 (General Electric Healthcare Life Sciences Division);

[0032] Porcine parvovirus: HW-1 strain;

[0033] Cell growth medium: DMEM containing 8% calf serum (Beijing Qingda Tianyi Biotechnology Co., Ltd.);

[0034] Virus maintenance solution: DMEM containing 1% calf serum by volume (Beijing Qingda Tianyi Biotechnology Co., Ltd.);

[0035] Cell culture: In 14L bioreactors, Cytodex-1 was added at a concentration of 10g / L. After hydration, eluted twice with pH 7.2 phosphate buffered saline PBS, sterilized, and inoculated with PK-15 cells. Carry out culture; the method parameters of culture are: pH7.2, temperature 37 ℃, DO50%, stirring speed 30~100rpm; take samples regularly every day to observe the growth of the cells, perform cell counts, and determine the consumption of glucose. When the density of the cells reaches 1.5× 10 6 perml, start perfusion, the rate of p...

Embodiment 3

[0040] Bioreactor: 14L and 40L bioreactors from NBS Company of the United States;

[0041] Microcarrier: Cytodex-1 (General Electric Healthcare Life Sciences Division);

[0042] Porcine parvovirus: HW-1 strain;

[0043] Cell growth medium: DMEM / F12 containing 8% calf serum by volume;

[0044] Virus maintenance solution: DMEM / F12 containing 1% calf serum by volume;

[0045] Cell culture: In 14L bioreactors, add Cytodex1 at a concentration of 10g / L, after hydration, wash with pH7.2 phosphate buffered saline PBS twice, sterilize, inoculate ST cells, and culture; The parameters of the method are: pH7.2, temperature 37°C, DO50%, stirring speed 30-100rpm; samples are taken regularly every day to observe the growth of the cells, count the cells, and measure the consumption of glucose. When the density of the cells reaches 1-1.5×10 6 / ml, start perfusion, and the perfusion speed depends on the cell density and glucose consumption at 0.5 to 4 working volumes per day, culture for 4 d...

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Abstract

The invention provides a method for industrially producing a swine parvovirus vaccine by utilizing a bioreactor, comprising the following steps of: (1) sterilizing a micro-carrier and the bioreactor, adding a cell growth solution, inoculating, preparing the vaccine and culturing with cells, inoculating a swine parvovirus after the cell on the micro-carrier forms a compact single layer, and continuously culturing to propagate the virus; (2) stopping culturing until the cytopathy reaches more than 80%, and harvesting a virus solution; (3) carrying out ultrafiltration concentration and virus inactivation on the harvested virus solution; and (4) purifying and inactivating the virus through a column chromatography method to prepare the vaccine. The invention has the advantages of favorable controllability of processing parameters, high cell density and virus titer, favorable vaccine safety, stable and reliable quality, high production efficiency, and the like.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a method for industrially producing porcine parvovirus vaccine using a bioreactor. Background technique [0002] Porcine parvovirus (PPV) is one of the main diseases causing reproductive disorders in sows. Pregnant sows mainly show abortion, stillbirth, mummified fetuses, weak piglets, etc., and the first-born sows are the most serious, causing huge economic losses to the global pig industry. At present, the prevention and control of porcine parvovirus is mainly based on immune prevention. Due to the single serotype of PPV and its high immunogenicity, vaccination has become an effective method to control the infection of porcine parvovirus. [0003] The vaccine widely used in my country is inactivated porcine parvovirus vaccine at present. The production of the vaccine adopts the traditional spinning bottle process. This process has been used in China's va...

Claims

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Application Information

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IPC IPC(8): A61K39/23C12N7/00A61P31/20C12R1/93
Inventor 秦红刚刘汉平漆世华李伟温文生李晶梅朱薇靖志强
Owner WUHAN CHOPPER BIOLOGY
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