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Method for producing swine pseudorabies live vaccine by using passage cell source, and product thereof

A technology for subculture cells and pseudorabies, applied in biochemical equipment and methods, medical preparations containing active ingredients, microorganisms, etc., can solve the lack of freezing and cold storage facilities in grassroots epidemic prevention departments, long-term vaccine storage and long-distance transportation restrictions, epidemic diseases epidemic and other problems, to achieve good immune protection, improve vaccine production and quality, and high immune efficacy

Active Publication Date: 2015-02-25
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the development of domestic traditional animal vaccine protective agents is relatively lagging behind, which greatly restricts the long-term storage and long-distance transportation of vaccines. In addition, the grassroots epidemic prevention departments lack necessary freezing and cold storage facilities, which may easily cause epidemics due to immunization failure.

Method used

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  • Method for producing swine pseudorabies live vaccine by using passage cell source, and product thereof
  • Method for producing swine pseudorabies live vaccine by using passage cell source, and product thereof
  • Method for producing swine pseudorabies live vaccine by using passage cell source, and product thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1 Preparation of heat-resistant protective agent

[0048] The composition and component content of the heat-resistant protective agent are shown in Table 1.

[0049] Table 1 Composition and content of heat-resistant protective agent

[0050]

[0051] For each heat-resistant protectant, 1 L of water for injection was heated to a temperature of 60° C. under agitation, and then the compound was slowly added to hot water while maintaining agitation via a magnetic bar to facilitate its dissolution. Agitation was maintained for about 20 minutes after the last addition, resulting in a homogeneous solution.

[0052] Then the solution was cooled to room temperature, and after cooling, the heat-resistant protective agent was sterilized by filtration through a filter membrane with a pore size of 0.22 μm, and stored at room temperature.

Embodiment 2

[0053] Example 2 Production process of porcine pseudorabies live vaccine produced by passage cells

[0054] The production process of producing porcine pseudorabies live vaccine with passaged cells, the concrete steps are as follows:

[0055] (1) Select passage cells as cells for seedling production.

[0056] (2) Subculture and culture of seedling cells: the above-mentioned subcultured cells were digested and subcultured with trypsin cell dispersion, and the cells containing 90% to 97% by volume of MEM culture medium and 3% to 10% by volume of fetal bovine serum were used for passage. The growth medium (pH adjusted to 7.0-8.0) was further cultured at 36°C-38°C to form a good monolayer, which was used for continued passage or virus inoculation.

[0057] (3) Propagation of cytotoxic species: Inoculate the well-grown monolayer of the above passaged cells with the attenuated strain of porcine pseudorabies virus, and use MEM culture medium containing 95% to 99% by volume and fetal...

Embodiment 3

[0060] Example 3 Preparation of virus liquid

[0061] Select porcine testis (ST) passage cells, pig kidney (PK15) passage cells and pig kidney (IBRS-2) passage cells, the attenuated strain of porcine pseudorabies virus is selected from porcine pseudorabies virus strain SA215, and the production process according to Example 2 is respectively Porcine pseudorabies virus liquid B1, B2 and B3 were prepared, and the titer was measured, and the virus content per 1ml was respectively 10 9.8 TCID 50 、10 9.75 TCID 50 、10 9.8 TCID 50 , stored above 0°C.

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Abstract

The present invention provides a method for producing a swine pseudorabies live vaccine by using a passage cell source, and a swine pseudorabies live vaccine product thereof. The method comprises: adopting passage cells to culture a swine pseudorabies virus attenuated strain; harvesting to obtain a cell culture virus solution; and adding a stabilizer, and carrying out freezing vacuum drying to obtain the passage cell derived swine pseudorabies live vaccine. During the use, the diluent of the prepared vaccine of the present invention can further be used. According to the present invention, the method has advantages of simple and stable production process, easy operation, high virus content, low difference between batches, controllable quality, significant vaccine yield and quality improving, and the like, the obtained product can be stored for a long time at the room temperature, especially at the temperature of 2-8 DEG C, the shelf life can be 36 months, and the diluted vaccine obtained by diluting the vaccine with the diluent of the present invention can be stored for a long time at the room temperature and can maintain the stability.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and more specifically relates to a method for producing porcine pseudorabies live vaccines with subcultured cell sources and products thereof. Background technique [0002] Pseudorabies is an acute infectious disease caused by pseudorabies virus (PRV) in many animals. The epidemic spreads throughout the world's major pig-growing countries. Immunization is the main strategy to control porcine pseudorabies. At present, the cells used in the production of porcine pseudorabies live vaccines in my country are chicken embryo fibroblast primary cells, resulting in large batch-to-batch differences and low toxin-producing titers, which brings difficulties to the improvement of vaccine production and efficacy. There are many miscellaneous proteins, which can easily cause allergic reactions. And the storage of semi-finished virus liquid needs to be stored below -15°C to avoid the re...

Claims

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Application Information

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IPC IPC(8): A61K39/245A61P31/22C12N7/00
Inventor 张许科孙进忠田克恭
Owner PU LIKE BIO ENG
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