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Method for preparing Seneca valley virus by utilizing suspension cell line

A technology of suspension cells and cell lines, applied in the field of veterinary biological products, can solve problems such as difficult control and no vaccine, and achieve the effect of reducing batch-to-batch differences and improving production capacity

Active Publication Date: 2019-12-31
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because there is no commercial vaccine for the time being, once the SVA epidemic occurs, it is difficult to effectively control it, which has become a problem that needs to be solved urgently in the world

Method used

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  • Method for preparing Seneca valley virus by utilizing suspension cell line
  • Method for preparing Seneca valley virus by utilizing suspension cell line
  • Method for preparing Seneca valley virus by utilizing suspension cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 acclimation of non-suspended BHK-21 cell line

[0045] Materials used in this example: adherent BHK-21 cell line, ATCC number: CCL-10; basal medium: Gibco MEM basic, serum-free medium: Changzhou Emines Biotechnology Co., Ltd.; virus: Sene Cardiac virus FJ / 001 strain (SVV / FJ / 001 strain) was isolated and preserved in our laboratory, and its microbial deposit number is: CCTCC NO: V201802.

[0046] First, the adherent BHK-21 cell line was acclimated to the suspension BHK-21-S cell line. The specific steps are as follows:

[0047] 1. Take out one preserved non-suspended BHK-21[C-13] cell line from the liquid nitrogen tank, and quickly place it in a 37°C water bath for thawing, then transfer it to a 15ml centrifuge tube and add 5-10ml 10% FBS MEM medium, 1000 rpm, centrifuge for 5 min, discard the supernatant, and gently pipette the cell pellet with 10 ml of complete medium containing 10% FBS MEM, then place it in a Corning T25 culture flask and place it at 37°C, c...

Embodiment 3

[0063] Example 3 Determination of serum-free medium for suspension BHK-21-S cell line

[0064] In order to eliminate the influence of serum in the low serum medium on subsequent virus culture, and at the same time reduce costs for industrialized production, the present invention selects a variety of serum-free medium to culture suspension BHK-21-S cell line. The medium screening alignment is as follows.

[0065]

[0066] Finally, the medium of Changzhou Emines Biotechnology Co., Ltd. is preferred, and the initial density of cell culture is 0.8-1.5×10 6 cell / ml, the cell density reaches 7.0×10 in 72h 6 cell / ml, culture temperature is 35-37°C, culture speed is 100-110rpm, CO 2 The concentration is 5%, and the culture shaker is preferably a box-type temperature-controlled shaker of Adolph Conai ISF1-X.

Embodiment 4

[0067] Example 4 Preparation of Seneca virus using suspension BHK-21-S cell line

[0068] Materials used in this example: Serum-free medium: purchased from Changzhou Aminens Biotechnology Co., Ltd.; Virus: Seneca virus is FJ / 001 strain (SVV / FJ / 001 strain), which was isolated and preserved in this laboratory , its microbial deposit number is: CCTCC NO: V201802; TPCK trypsin from bovine pancreas, purchased from sigma company, item number: 9002-07-7; air-permeable conical flask: Corning 125ml.

[0069] 4.1 Determination of the most suitable pH for exposure

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Abstract

The invention provides a method for preparing Seneca valley virus by utilizing a suspension cell line. The method for preparing the Seneca valley virus by utilizing the suspension cell line comprisesthe following steps: (1) domesticating a non-suspension BHK-21 cell line into a suspension BHK-21-S cell line; and (2) culturing the Seneca valley virus by using the suspension BHK-21-S cell line obtained in the step (1). By culturing the Seneca valley virus by using the suspension BHK-21-S cell line obtained by performing domestication, yield of the Seneca valley virus can be effectively increased; and thus, foundation is laid for large-scale production of vaccines to the Seneca valley virus.

Description

technical field [0001] The invention relates to a method for preparing Seneca virus by utilizing a suspension cell line, in particular to a method for preparing Seneca virus by utilizing a suspension culture passage cell line, and belongs to the technical field of veterinary biological products. Background technique [0002] Seneca virus A (Seneca virus A, SVA), also known as Seneca valley virus (SVV), is a new and newly introduced virus in my country. It is a new Seneca virus in the Picornaviridae family. The only member of the virus genus, the clinical symptoms are indistinguishable from foot-and-mouth disease. Before 2014, SVA only occurred sporadically in the United States and Canada, but since 2015, SVA epidemics have occurred in Brazil, the United States, Thailand, and China, and have continued to spread, increasing the harm. Since SVA was introduced into my country in 2015, epidemics have occurred in Guangdong, Hubei, Heilongjiang, Fujian, Guangxi, Henan, Hebei, Shand...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/02C12N7/00
CPCC12N5/0686C12N7/00C12N2770/32051
Inventor 郑海学曹伟军杨帆朱紫祥蒋保余魏婷郑敏田宏张克山刘永杰茹毅党文刘湘涛
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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