Preparation method of mink canine distemper virus live vaccine and vaccine prepared by same

A canine distemper virus and live vaccine technology, applied in biochemical equipment and methods, vaccines, viruses, etc., can solve the problems of uncontrollable environment for culturing cells, difficult to expand production, low degree of automation, etc., and achieve a simple and stable production process. , The effect of improving production efficiency and high degree of automation

Inactive Publication Date: 2016-08-03
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many disadvantages in the traditional spinning bottle process, such as: low degree of automation, high labor intensity; uncontrollable environment for culturing cells, easy to be polluted by the environment; long time-consuming, low efficiency, high production cost, difficult to expand production; large quality differences between different batches ; involving biosafety and public health issues

Method used

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  • Preparation method of mink canine distemper virus live vaccine and vaccine prepared by same
  • Preparation method of mink canine distemper virus live vaccine and vaccine prepared by same
  • Preparation method of mink canine distemper virus live vaccine and vaccine prepared by same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] A method for the preparation of mink distemper virus live vaccine is provided in Embodiment 1 of the present invention, comprising the following steps:

[0059] Step 101: The Marc-145 cells used for seedling production were divided into 4×10 5 Individual / mL density is inoculated in the bioreactor of 5L and is cultivated with microcarrier; The microcarrier that is used for cultivating is Cytodex microcarrier, and its use density is 3g / L; The cell culture medium is: DMEM culture medium containing 5% newborn bovine serum; the culture conditions when using microcarriers are: temperature 36°C, CO 2 The content is 4.8%, the stirring speed is 55rpm, the dissolved oxygen is 45%, the pH is 7.2, and the reactor is automatically controlled for cultivation;

[0060] Step 102: After the sensitive cells are cultured for 48 hours, more than 70% of them grow into a dense monolayer, and the canine distemper virus CDV3-CL strain is inoculated into the bioreactor according to the ratio o...

Embodiment 2

[0063] A method for the preparation of mink canine distemper virus live vaccine is provided in Embodiment 2 of the present invention, comprising the following steps:

[0064] Step 201: Divide the MDCK cells for seedling production into 2×10 6 Individual / mL density is inoculated in the bioreactor of 5L and is cultivated with microcarrier; The microcarrier that is used for cultivating is Cytodex microcarrier, and its use density is 10g / L; The cell culture medium is: DMEM culture medium containing 8% newborn bovine serum; the culture conditions when cultured with microcarriers are: temperature 38°C, CO 2 The content is 5.2%, the stirring speed is 65rpm, the dissolved oxygen is 55%, the pH is 7.4, and the reactor is automatically controlled for cultivation; the method of culturing cells is batch cultivation;

[0065] Step 202: After the sensitive cells described in step 201 are cultured for 20 hours, more than 50% of them grow into a dense monolayer, and the canine distemper viru...

Embodiment 3

[0068] In order to describe the technical solution of the present application in more detail, the present invention also obtains Embodiment 3 through further refinement and limitation of each operation on the basis of Embodiments 1 and 2, including the following steps, please refer to figure 1 :

[0069] Equipment and reagents used:

[0070] Bioreactor: Qizhi Bioengineering Equipment Co., Ltd. 14L bioreactor;

[0071] Microcarrier: GE Cytodex-1;

[0072] Canine distemper virus for seedling production: CDV3-CL strain, identified, kept and supplied by the Institute of Special Products of the Chinese Academy of Agricultural Sciences;

[0073] DMEM medium (dry powder): GIBCO company;

[0074] Newborn bovine serum: Inner Mongolia Jinyuankang Bioengineering Co., Ltd.;

[0075] Trypsin (dry powder): GIBCO company.

[0076] Step 301: Preparation of bioreactor and microcarrier

[0077] Thoroughly clean the tank body of the bioreactor, connect the inlet and outlet pipelines and ga...

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Abstract

The invention relates to the field of veterinary biological products and particularly relates to a preparation method of a mink canine distemper virus live vaccine. The method comprises the following steps: inoculating a bioreactor with sensitive cells for vaccine preparation, and culturing by using a micro-carrier; after the sensitive cells are cultured by over 50% and grow into a dense single layer, inoculating the bioreactor with canine distemper virus for enrichment culture; harvesting the virus culture liquid and micro-carrier; performing freeze-thawing and removing the micro-carrier and cell debris to obtain virus liquid; and blending the virus liquid to obtain the vaccine. By improving the reaction conditions of each step and optimizing the production flow, the method provided by the invention realizes the technical effects of short production cycle, high virus titer, stable product quality, increased production efficiency and low side effect.

Description

technical field [0001] The invention relates to the field of veterinary biological products, in particular to a method for preparing mink canine distemper virus live vaccine and the vaccine prepared by the method. Background technique [0002] Mink distemper, also known as mink distemper, is an acute, febrile and highly infectious highly contagious infection caused by Canine distempervirus (CDV) of the family Paramyxoviridae and the genus Morbillivirus. Sexually transmitted diseases are one of the main infectious diseases in mink farming. [0003] Reactor suspension culture technology has been widely used in the production of biopharmaceuticals in the world, and has continued to develop in the construction of high-expression cell lines and the development of personalized culture media. A large number of new reactors and the increase in product expression have led to increased reactor capacity. Oversupply, the market in developed countries is saturated and expanding to devel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12C12N7/00C12N5/00C12N5/02A61P31/14
CPCA61K39/12C12N5/0075C12N7/00A61K2039/525A61K2039/552C12N2531/00C12N2760/18451C12N2760/18434
Inventor 尹茉莉冯二凯任飞程世鹏陈立志彭风华盛程程王萃瑜王振军李云松
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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