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Method for producing porcine pseudorabies virus antigen by full suspension cell culture

A porcine pseudorabies virus and cell culture technology, applied in the field of full suspension cell culture to produce porcine pseudorabies virus antigen, achieves the effects of small batch-to-batch difference, stable quality, simple operation and easy expansion

Pending Publication Date: 2019-09-17
JIANGSU NANNONG HI TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the deficiencies of the existing production and culture methods, and provides a method for the production of porcine pseudorabies virus antigen by whole suspension cell culture

Method used

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  • Method for producing porcine pseudorabies virus antigen by full suspension cell culture
  • Method for producing porcine pseudorabies virus antigen by full suspension cell culture
  • Method for producing porcine pseudorabies virus antigen by full suspension cell culture

Examples

Experimental program
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Effect test

Embodiment 1

[0029] ——Production of porcine pseudorabies virus antigen by whole suspension cell culture

[0030] 1. Preparation of porcine pseudorabies virus antigen

[0031] (1) Cultivation and passage of seed cells

[0032] Seed cells PK-15B1 (CCTCC No.C200936, provided by Professor Jiang Ping of Nanjing Agricultural University) were cultured and grown in shake flasks, samples were taken for cell counting, and the cell density reached 3.7×10 6 cells / ml, after adding serum-free cell culture medium CDPK15259, according to 0.5×10 6 The density of each / ml was added to the reactor for culture, and the culture volume was half of the final culture medium volume of the reactor, 2.5L;

[0033] (2) Inoculation of seed cells

[0034] The measured cell density reached 6.4×10 6 Individual / ml, the PRVZJ011G (CGMCC No.7957) kind of venom of 5% of the final culture solution volume of the reactor is inserted into the seed cells, and the PRVZJ011G kind of venom virus content is 10 7.0 TCID 50 / ml, an...

Embodiment 2-4

[0051] Examples 2-4 are based on Example 1, using different formulations of serum-free cell culture medium CD PK15259 for cultivation (the process parameters of each step are kept the same), and the formula of serum-free cell culture medium CD PK15 259 is shown in Table 1.

Embodiment 2

[0054] The porcine pseudorabies virus antigen semi-finished product virus content of embodiment 2 10 9.33 TCID 50 / mL;

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Abstract

The invention provides a method for producing porcine pseudorabies virus antigen by full suspension cell culture. The method is capable of greatly reducing production cost, free of carrier or serum, more simple and convenient in production process and free of cell habituation or liquid exchange operation, and the technology is small in area occupation, capable of rapidly carrying out large-scale production, relatively low in environmental requirements, high in automation degree, low in labor cost, small in difference between production batches, stable and easy-control in quality and capable of obviously improving the yield and quality of products. The method includes the following steps: (1) culture and passage of seed cells (PK-15B1); (2) inoculation of seed cells (seed virus, PRV-ZJ011G); and (3) harvesting and content determination of virus; and serum-free cell culture medium CD PK15 259 is used in each step.

Description

technical field [0001] The invention relates to a method for producing porcine pseudorabies virus antigen by whole suspension cell culture, belonging to the field of veterinary biological products. Background technique [0002] Porcine pseudorabies virus (Pseudorabies virus, PRV) is a member of the herpesviridae herpesvirus A subfamily. It can cause central nervous system disorders in newborn piglets, respiratory symptoms in growing pigs, and can cause abortion, stillbirth and weak piglets in pregnant sows. The United States and some European countries have declared the eradication and eradication of the disease. The gene-deficient attenuated vaccine is widely used in my country to effectively control and gradually purify it. However, as pseudorabies has mutated in recent years and the virulence has returned to strength, the protective effect of traditional strain vaccines has declined significantly. Therefore, the market urgently needs pseudorabies vaccines of mutated stra...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N5/071C12R1/93
CPCC12N7/00C12N5/0686C12N2710/16751C12N2500/90
Inventor 董彦鹏肖澄钱建宁罗顺姜平缪芬芳张业忻车巧林王嘉琪胡芳沈燕胡静雅王彦奇柴玮迹耿晓眉
Owner JIANGSU NANNONG HI TECH