A kind of method that utilizes suspension cell line to prepare Seneca virus

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: 2020-10-16
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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  • A kind of method that utilizes suspension cell line to prepare Seneca virus
  • A kind of method that utilizes suspension cell line to prepare Seneca virus
  • A kind of method that utilizes suspension cell line to prepare Seneca virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Domestication of non-suspension 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 Amines Biotechnology Co., Ltd.; virus: Sene Card virus FJ / 001 strain (SVV / FJ / 001 strain), isolated and preserved by this laboratory, its microbial deposit number is: CCTCC NO:V201802.

[0046] First, the adherent BHK-21 cell line was domesticated into a suspension BHK-21-S cell line, the specific steps are as follows:

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

Embodiment 3

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

[0064] In order to eliminate the influence of serum in the low-serum culture medium on the subsequent virus culture and at the same time reduce the cost of industrialized production, the present invention screens a variety of serum-free culture media to cultivate the suspended BHK-21-S cell line. The media selection and comparison are as follows.

[0065]

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

Embodiment 4

[0067] Example 4 Preparation of Seneca virus using suspended 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 strain FJ / 001 (SVV / FJ / 001 strain), isolated and preserved in this laboratory , Its microbial preservation number is: CCTCC N0: V201802; TPCK Trypsin from bovine pancreas, purchased from sigma company, item number: 9002-07-7; breathable triangle shake flask: Corning 125ml.

[0069] 4.1 Determination of the most suitable pH

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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 using a suspension cell line, in particular to a method for preparing Seneca virus by using a suspension culture subculture 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 virus emerging from the world and newly introduced into my country. It is a new Seneca in the Picornavirus family. The only member of the virus genus, 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, China, etc., and they have continued to spread, increasing the damage. Since the introduction of SVA to my country in 2015, epidemics have occurred in Guangdong, Hubei, Heilongjiang, Fujian, Guangxi, Hen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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