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Method for manufacturing Marek's disease vaccine by utilizing cell factory

A technology for chicken Marek's disease and production method, which is applied in the directions of biochemical equipment and methods, microorganisms, antiviral agents, etc. The effect of stable immune effect, high immune effect, and easy operation

Active Publication Date: 2011-04-06
北京盛华四合生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Spinner bottle culture is currently a cell culture method commonly used by domestic vaccine manufacturers. It has the advantages of simple structure, less investment, mature technology, good repeatability, and easy expansion. At the same time, there are also low cell growth densities and differences in cell growth between spinner bottles. Disadvantages such as large size, high labor intensity, large space occupation, high energy consumption, and large water consumption

Method used

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  • Method for manufacturing Marek's disease vaccine by utilizing cell factory
  • Method for manufacturing Marek's disease vaccine by utilizing cell factory
  • Method for manufacturing Marek's disease vaccine by utilizing cell factory

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Choose Cell STACK 10 layers (636cm) produced by American CORNING Company 2 / layer), the preparation process of chicken Marek's disease liquid nitrogen seedlings prepared by synchronous infection method.

[0037] The main steps of liquid nitrogen vaccine preparation process are as follows:

[0038] (1) Preparation of chicken embryo fibroblast monolayer

[0039] Select 10-day-old well-developed SPF chicken embryos, first disinfect the air chamber with 4% iodine, deiodine with 75% alcohol, take out the chicken embryos aseptically, wash the embryo body with Hank's solution, and cut the embryo body with sterile scissors. Tissue pieces the size of rice grains were washed with Hank's solution for 3 times; 0.25% trypsin (DE) was added at a rate of 4 mL per chicken embryo, digested at 38°C for 30 minutes, the trypsin was discarded, and washed with Hank's solution for 3 times; add 2mL of nutrient solution to each chicken embryo to make a cell suspension containing 3 million vi...

Embodiment 2

[0055] Choose Cell STACK 40 layers (636cm) produced by American CORNING Company 2 / layer), the preparation process of chicken Marek's disease liquid nitrogen seedlings prepared by single-layer infection method.

[0056] The main steps of liquid nitrogen vaccine preparation process are as follows:

[0057] (1) Preparation of chicken embryo fibroblast monolayer

[0058] Select 9-day-old well-developed SPF chicken embryos, first disinfect the air chamber with 4% iodine, deiodine with 75% alcohol, aseptically take out the chicken embryos, remove the head and viscera, put them in a sterile glass container, and use Wash the embryo body with Hank's solution, cut the embryo body into rice grain-sized tissue pieces with sterile scissors, wash with Hank's solution three times, add 0.25% trypsin at a rate of 6 mL per chicken embryo, digest at 38°C for 20 min, Discard the trypsin, wash with Hank's solution 3 times; add 4 mL of nutrient solution to each chicken embryo to make a cell susp...

Embodiment 3

[0074] An experiment was designed to compare the curative effect of the chicken Marek's disease liquid nitrogen vaccine (814 strains) produced in Example 1 and the CVI988 vaccine produced by 2 foreign manufacturers to prevent chicken Marek's disease.

[0075] 1. A total of 132 1-year-old SPF Bailaihang chicks (provided by Beijing Merial Weitong Experimental Technology Co., Ltd.) were randomly divided into 6 groups, 22 in each group. The immunization group consisted of 4 groups, which were inoculated with 814 low-generation virus vaccine, 814 strain vaccine, CVI988-A, and CVI988-B respectively, and 3000 PFU was injected subcutaneously at the back of each mouse. Non-immune group 2, after immunization, were isolated and raised in the isolator for 21 days. 21 days after immunization, 22 mice in the immunization group and the positive control group were inoculated with the standard virulent strain of Marek's disease (MD) Jing-1 strain (purchased from: Institute of Animal Husbandry ...

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Abstract

The invention belongs to the technical field of biological medicament making, and discloses a method for preparing a Marek's disease liquid nitrogen vaccine (814-strain) by utilizing a cell factory and the Marek's disease liquid nitrogen vaccine (814-strain) prepared by the method. The Marek's disease liquid nitrogen vaccine (814-strain) produced by utilizing the cell factory has high quality, a high immune efficiency and a stable immune effect; in addition, the Marek's disease liquid nitrogen vaccine also has the advantages of low generation, small plaque, high plaque forming unit (PFU) content, high immunogenicity, low cost, capacity of generating immunoprotection in a relatively short time and the like.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, in particular to a method for producing chicken Marek's disease liquid nitrogen vaccine (814 strains), and in particular discloses a method for preparing chicken Marek's disease liquid nitrogen vaccine (814 strains) by using a cell factory . Background technique [0002] Chicken Marek's disease is a highly contagious lymphoproliferative disease of chickens caused by cell-associated herpesvirus (MDV). Chicken flocks infected with Marek's virus are more harmful and cause serious harm to the poultry industry. With the widespread application of vaccines and the continuous selection of chicken strains, MDV has continuously increased its virulence under the dual effects of natural selection and immune pressure, breaking through the protection provided by vaccines time and time again. [0003] In the 1970s, the standard virulent virus (VMDV) was isolated. From the end of the 1970s to the 198...

Claims

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Application Information

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IPC IPC(8): A61K39/255C12N7/00A61P31/22
Inventor 马力王文泉张连祥朱秀芝陈玲周海琴金剑李静袁芳郎立群张洪郑杰贾桂珍李然曹宁车竞常如军张发明张红邢雪莲
Owner 北京盛华四合生物科技有限公司
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