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Production method of canine parvovirus inactivated vaccine by utilizing bioreactor

A bioreactor, canine parvovirus technology, applied in antiviral agents, pharmaceutical formulations, medical preparations containing active ingredients, etc., can solve the problems of uneven and stable product quality, large batch-to-batch variation, and low production efficiency.

Inactive Publication Date: 2013-01-09
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Vaccination of animals is the most effective way to prevent canine parvovirus infection. At present, the methods of domestic production of canine parvovirus vaccine mostly use the traditional spinner bottle method or chicken embryo production process, but it is not suitable for the current large-scale animal vaccine production. It is reflected in the low titer of cultured virus, uneven and stable product quality, large batch-to-batch variation, and low production efficiency, etc.

Method used

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  • Production method of canine parvovirus inactivated vaccine by utilizing bioreactor
  • Production method of canine parvovirus inactivated vaccine by utilizing bioreactor
  • Production method of canine parvovirus inactivated vaccine by utilizing bioreactor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 10L AMP rapid flow bioreactor paper carrier system culture

[0033] The bioreactor is an AMP AP20 torrent bioreactor equipped with a disposable polyester fiber paper carrier system. The cells are cat kidney passage cells F81, which were purchased from Shanghai Yanjing Biochemical Co., Ltd., and the canine parvovirus used for seedling production is CPV-2 strain, and its ATCC preservation number is VR-2016. DMEM / F12 medium was purchased from Beijing Tsingda Tianyi Company, and calf serum was purchased from Wuhan Sanli Biotechnology Co., Ltd.

[0034] 1) Preparation of bioreactor and processing of paper carrier

[0035] Aseptically install the electrode in the cell culture bag and pump in fresh medium.

[0036]The cell growth medium was DMEM / F12 liquid medium supplemented with 10% calf serum.

[0037] Introduce sterile pH 7.2 phosphate-buffered saline (PBS) into the perfusion culture bag pre-loaded with paper carrier and soak overnight. The PBS with pH 7.2 wa...

Embodiment 2

[0053] Embodiment 2 14L NBS stirring type bioreactor scale-up cultivation

[0054] The reactor is a Celligen 310 bioreactor from NBS Company with a Cell-Lift stirring paddle. The microcarrier is the Cytodex1 carrier of GE Company, the cell growth medium is DMEM / F12 medium (Beijing Tsingda Tianyi Biotechnology Co., Ltd.) with 10% calf serum (Wuhan Sanli Biotechnology Co., Ltd.), and the cat kidney passage cells F81 was purchased from Shanghai Yanjing Biochemical, and the CPV-2 strain was the ATCC (American Type Culture Collection) VR-2016 strain.

[0055] 1) Preparation of the bioreactor

[0056] Thoroughly clean the tank body of the bioreactor, calibrate the pH and DO electrodes and install them on the tank body, bandage the pipe joints and conduct a leak test. After completion, perform high-pressure steam sterilization. Naturally cool to room temperature. The aseptically filtered cell growth medium is aseptically connected to the reactor, and introduced into the tank throu...

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Abstract

The invention discloses a production method of a canine parvovirus inactivated vaccine by utilizing a bioreactor. The production method comprises the following steps: (1) culturing a vaccine producing cell by applying a microcarrier and chip carrier system of the bioreactor; (2) vaccinating a canine parvovirus and performing virus multiplication culture; (3) harvesting virus culture fluid; and (4) inactivating the virus fluid with BEI (binary ethyleneimine), and adding adjuvant to prepare the vaccine. The canine parvovirus inactivated vaccine has the advantages of high density of cultured cell, high virus titer, uniform and stable quality, controllable process and high production efficiency, and the defects of large difference among product batches, low antigen content and low production efficiency in a conventional spinner bottle or chick embryo production process can be overcome.

Description

technical field [0001] The invention belongs to the field of veterinary biological products. More specifically, the present invention relates to a method for industrial production of viral vaccines using bioreactors. Background technique [0002] Canine parvovirus disease (Canineparvovirus disease, CPVD) is an acute infectious disease of dogs caused by canine parvovirus (Canineparvovirus, CPV). It can be transmitted between dogs and cats. Dogs of all ages can be infected. Young dogs after weaning are the most susceptible. The transmission speed is fast, and the morbidity and mortality rate are as high as 70%. Since the official report of the prevalence of canine parvovirus in my country, the disease has caused great harm to the dog industry in my country. Vaccination of animals is the most effective way to prevent canine parvovirus infection. At present, the methods of domestic production of canine parvovirus vaccine mostly use the traditional spinning bottle method or chi...

Claims

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Application Information

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IPC IPC(8): A61K39/23A61P31/20
Inventor 李伟刘洁秦红刚漆世华朱薇谢红玲韩兴王桢桢王威温文生
Owner WUHAN CHOPPER BIOLOGY
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