Method for preparing duck hemorrhagic ovaritis inactivated vaccines

A technology of hemorrhagic oophoritis and inactivated vaccines, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., to achieve the effects of improving sensitivity, saving manpower, and increasing the number of cells

Active Publication Date: 2013-06-19
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are domestic research reports on the preparation of duck hemorrhagic oophoritis vaccine by usi

Method used

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  • Method for preparing duck hemorrhagic ovaritis inactivated vaccines
  • Method for preparing duck hemorrhagic ovaritis inactivated vaccines
  • Method for preparing duck hemorrhagic ovaritis inactivated vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Cloning, purification and screening of cells for seedling production by limiting dilution method (hamster kidney cells BHK21)

[0043] Young hamster kidney cells BHK21 were purchased from China Veterinary Drug Administration.

[0044]The duck hemorrhagic oophoritis virus vaccine strain is the duck hemorrhagic oophoritis virus HB strain, which was preserved in the China Center for Type Culture Collection on July 1, 2011, with the preservation number CCTCC V201122; the preservation address is: Wuhan University, China. The virus is an ssRNA virus with an envelope; the virus particles are roughly spherical, with a diameter of 40-60nm; the virus has strong resistance to the external environment, and it can be stored at 4°C for several weeks and stored at -20°C for several months , its infectivity is not affected; the virus is sensitive to ether and chloroform; most detergents can quickly inactivate it; at 37°C, it can be inactivated by fumigation with 0.1% formalin...

Embodiment 2

[0069] Example 2 Preparation of Duck Hemorrhagic Ovariitis Inactivated Vaccine by AP20C Torrent Perfusion Bioreactor

[0070] In the AP20C torrent perfusion bioreactor used in this example, the perfusion bag 6 has a volume of 8 L (both perfusion bags have a volume of 4 L), the torrent tank 5 has a volume of 13 L, and the perfusion bag 6 contains 300 g of polyester fiber paper Tablet carrier 8 (two perfusion bags each contain 150g carrier); the theoretical effective culture volume is 21L.

[0071] The cell is the same as the hamster kidney cell BHK21 preserved by the strain in Example 1;

[0072] Seed virus is the same as embodiment 1, a certain batch of duck hemorrhagic oophoritis virus HB strain seed virus, virus titer is 10 7.7 TCID 50 / ml.

[0073] The formula of the cell growth solution is 92% high-sugar DMEM solution by volume, 8% newborn bovine serum, and the pH value is adjusted to 7.3;

[0074] The cell maintenance solution is 98% high-sugar DMEM solution by volume...

Embodiment 3

[0104] Example 3 Preparation of duck hemorrhagic ovariitis inactivated vaccine by AP200 torrent perfusion bioreactor

[0105] The torrent perfusion bioreactors used in this example include AP20C type and AP200 type.

[0106] The AP20C type torrent perfusion bioreactor is the same as in Example 2.

[0107] Seedlings are all the same as in Example 2 with cells and seed poisons.

[0108] The AP200 perfusion bag 6 has a volume of 80L (the volume of the two perfusion bags is 40L), the surge tank 5 has a volume of 130L, and the perfusion bag 6 contains 3000g of polyester fiber paper carrier 8 (each of the two perfusion bags contains 1500g of carrier); Theoretical effective culture volume is 210L.

[0109] A. Preparation:

[0110] Two torrent perfusion bioreactors, AP20C and AP200, were selected to prepare for cell culture.

[0111] B, preparation work, concrete steps are as follows:

[0112] a. Cultivation of cells for seedling production

[0113] According to the method of Ex...

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Abstract

The invention relates to a method for preparing duck hemorrhagic ovaritis inactivated vaccines, and belongs to the field of biotechnology. The method for preparing the duck hemorrhagic ovaritis inactivated vaccines includes the following steps: (1) culturing of cells used for vaccine preparation, (2) virus inoculation and culture, (3) virus liquid harvest, (4) virus liquid inactivation, and (5) vaccine preparation. Cells used for vaccine preparation are screened, matching degree between the cells and the virus is strengthened, a riptide perfusion type bioreactor culture system is used for improving multiplication titer and harvest yield of the virus, and a whole production process does not involve other biosafety and public health problems and is suitable for large scale production.

Description

technical field [0001] The invention relates to a method for preparing an inactivated vaccine of duck hemorrhagic oophoritis by using a torrent perfusion bioreactor, belonging to the field of biotechnology Background technique [0002] Duck Hemorrhagic Ovaritis (DHO) is an acute, highly contagious duck infectious disease caused by infection with Duck Hemorrhagic Ovaritis Virus (DHOV), a member of the Flaviviridae family , can cause a sharp drop in feed intake and egg production rate of ducks of different ages, and even paralysis. Dissection mainly shows ovarian hemorrhage and atrophy, follicle deformation and degeneration, yolk, testis, fallopian tube and vas deferens atrophy. [0003] In 2010, duck hemorrhagic oophoritis disease was first discovered in Zhejiang, China, and then occurred in Jiangsu, Shandong, Hebei and Beijing. It is mainly seen in laying ducks. The incidence rate of infected ducks is almost 100%, the mortality rate is 5%-30%, and the egg production rate c...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61P31/14C12N7/00C12R1/93
Inventor 刘月焕何平有毛雅元刘涛郑朝朝韩春华林健徐倩倩杨保收梁武郁宏伟李建丽
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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