Method for producing duck tembusu virus inactivated vaccines in large scale

A technology for duck tembusu virus and inactivated vaccine, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, antiviral agents, etc., can solve the problem of low production efficiency, unsuitable for large-scale production of vaccines, and high labor intensity of operation And other issues

Inactive Publication Date: 2013-06-12
QILU ANIMAL HEALTH PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these raw materials and production methods have low operational technical requirements and mature and stable processes, they are labor-intensive in operation, low in production efficiency, and have large differences between vaccine batches, so they are not suitable for the large-scale production of current vaccines.

Method used

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  • Method for producing duck tembusu virus inactivated vaccines in large scale
  • Method for producing duck tembusu virus inactivated vaccines in large scale
  • Method for producing duck tembusu virus inactivated vaccines in large scale

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Preparation of Duck Tembusu Virus Inactivated Vaccine

[0040]Bioreactor: 14L and 40L stirred bioreactor, bioreactor setting parameters are: pH: 7.0, temperature: 36.5°C, dissolved oxygen: 50%, stirring speed: 60r / min;

[0041] Microcarrier: Cytodex1 (manufactured by GE)

[0042] Duck Tambusu virus: WFG36 strain, a strain of Duck Flavivirus (Duck Flavivirus) WFG36 strain, this virus strain has been sent to the Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing on April 12, 2011 Preserved by the General Microorganism Center of China Microbiological Culture Collection Management Committee, the number is: CGMCCNo.4718;

[0043] Cell growth medium: DMEM / F12 containing 10% fetal bovine serum (manufactured by GIBCO)

[0044] Virus maintenance solution: DMEM containing 5% fetal bovine serum (manufactured by GIBCO)

[0045] Cell culture:

[0046] (1) Add Cytodex-1 to a 14L bioreactor at a concentration of 15g / L...

Embodiment 2

[0056] Preparation of Duck Tembusu Virus Inactivated Vaccine

[0057] Bioreactor: 14L and 40L stirred bioreactor, bioreactor setting parameters are: pH: 7.0~7.4, temperature: 36.5~37℃, dissolved oxygen: 30~60%, stirring speed: 30~90r / min ;

[0058] Microcarrier: Cytodex1;

[0059] Duck Tembusu virus: WFG36 strain;

[0060] Cell growth medium: DMEM / F12 containing 10% fetal bovine serum ()

[0061] Virus maintenance solution: DMEM containing 5% fetal bovine serum (purchased from GIBCO)

[0062] Cell culture:

[0063] (1) Add Cytodex-1 to a 14L bioreactor at a concentration of 15g / L and sterilize;

[0064] (2) Add BHK cells for culture and observe every day;

[0065] (3) Wait until the cell density of the 14L bioreactor reaches 3.0×10 6 / ml, collect the microcarriers full of cells into a closed container and inoculate into a 40L bioreactor;

[0066] (4) Wait until the cell density of the 40L bioreactor reaches 2.5×10 6 / ml, inoculate duck Tembusu virus WFG36 strain, the ...

Embodiment 3

[0074] Inspection of finished product of duck hemorrhagic ovariitis inactivated vaccine

[0075] The vaccine tested in this example is prepared according to the technical scheme provided by the present invention.

[0076] [Properties] The appearance of the vaccines are all milky white emulsions.

[0077] Dosage form oil-in-water type. Take a clean straw, suck a small amount of vaccine and drop it in cold water, except for the first drop, it will not spread.

[0078] Stability Draw 10ml of vaccine into a centrifuge tube, centrifuge at 3000r / min for 15 minutes, no water phase precipitates out at the bottom of the tube.

[0079] Viscosity Use a 1ml straw (the inner diameter of the lower mouth is 1.2mm, and the inner diameter of the upper mouth is 2.7mm) to draw 1.0ml of vaccine at about 25°C, let it flow out vertically and naturally, and record the time required for the outflow of 0.4ml, all within 8 seconds.

[0080] [Sterility test] Carried out according to the method stipul...

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PUM

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Abstract

The invention discloses a method for producing duck tembusu virus inactivated vaccines in a large scale. According to the method, high-density culture of cells is performed by using a bioreactor culture technology to produce the duck tembusu virus inactivated vaccines. Compared with a conventional rotating bottle production process, the method for producing duck tembusu virus inactivated vaccines in a large scale has the characteristics that the automated control degree is high, and the production can be monitored in real time to ensure that the product quality is uniform and stable; and manpower and material resources can be saved during production to reduce the production cost; and the produced venom is high in virus content and low in inter-batch difference, and side effects are reduced.

Description

technical field [0001] The invention relates to a method for large-scale production of an inactivated duck Tembusu virus vaccine, which belongs to the technical field of veterinary biological products. Background technique [0002] Since April 2010, Fujian, Shandong, Zhejiang, Shanghai, Jiangsu and other places in my country's main laying duck breeding areas have successively broken out a disease characterized by a sudden drop in egg production of laying ducks and breeding ducks, and hemorrhagic oophoritis. Acute infectious disease characterized by lesions. In the early stage of the epidemic, according to the main lesions and clinical features, the epidemic was called duck hemorrhagic ovariitis (Duck Hemorrhagic Ovaritis, DHO). With the in-depth study of etiology, the disease was confirmed to be caused by a new type of flavivirus (Duck Flavivirus)—Duck Tembusu virus (DTMUV). In 2011, the first waterfowl epidemic prevention and control seminar of the Chinese Association of A...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61P31/14C12N7/00C12R1/93
Inventor 鲍海忠王蕾徐龙涛张青婵吴昊巩志宏李晓静彭建云张中波陈茹张丹
Owner QILU ANIMAL HEALTH PROD
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