Method for preparing vaccine by Newcastle disease virus cultured by using chick embryo continuous cell line and bioreactor

A technology of bioreactor and Newcastle disease virus, which is applied in the field of chicken embryo passage cell line and bioreactor to cultivate Newcastle disease virus to prepare vaccines, can solve the biological safety hazard of exogenous virus contamination from embryo body source, and the process of primary fibroblasts cumbersome, high degree of chicken heterogeneity, etc., to avoid greater stress, good economic benefits and application prospects, and increase the effect of virus titer

Active Publication Date: 2014-12-17
浙江美保龙生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The direct use of embryoid bodies for virus propagation is easily limited by the source of non-immune embryoid bodies. At the same time, the use of embryoid bodies for virus propagation will bring safety hazards to vaccine preparation due to the large amount of miscellaneous proteins in the cultured virus suspension, and improper disposal of embryoid body waste. There is also the risk of poisoning
Although the spinner bottle primary cell culture technology is relatively mature, the process of preparing primary fibroblasts is cumbersome, and there are large differences between batches. The amount of cultured virus is small and the toxicity is low. Security risks
[0004] The patent application with the patent publication number CN102600465A discloses a production method of chicken Newcastle disease live vaccine and its products. The cell lines used in the method for cultivating Newcastle disease virus in this patent include MDCK cell line, vero cell line, marc-145 cell line , ST cell lines, BHK cell lines or MDBK cell lines, and the method of cultivating Newcastle disease virus to prepare live vaccines using chicken embryo passage cell lines has not been reported
The above-mentioned cell lines have large interspecies differences for poultry, and the vaccines prepared from this have a high degree of heterogeneity for chickens, which is likely to cause greater irritation

Method used

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  • Method for preparing vaccine by Newcastle disease virus cultured by using chick embryo continuous cell line and bioreactor
  • Method for preparing vaccine by Newcastle disease virus cultured by using chick embryo continuous cell line and bioreactor

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Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1 Cultivate Newcastle disease virus and prepare live vaccine with chicken embryo passage cell line and bioreactor, comprise the following steps:

[0030] (1) Selection of bioreactor: Rapid flow reactor AP-20 with a volume of 7L.

[0031] (2) Selection of cells: chicken embryo cell line DF-1, a cell line suitable for the growth of chicken Newcastle disease virus, non-carcinogenic, purchased from the American Type Culture Collection;

[0032] (3) Selection of virus strains: the chicken Newcastle disease virus La sota strain was selected, which can be purchased from the China Veterinary Drug Control Institute.

[0033] (4) Cultivation of different cells: DF-1 cells are cultured in a square bottle, generally passaged at a ratio of 1:3 to 1:5, digested and counted after the cells have grown to a single layer, and used for large-scale cell culture.

[0034] (5) Propagation of cell-adapted virus species: take the cell-adapted Newcastle disease virus virus species, ...

Embodiment 2

[0045] Embodiment 2 Cultivate Newcastle disease virus and prepare inactivated vaccine with chicken embryo passage cell line and bioreactor, comprise the steps:

[0046] (1) Selection of bioreactor: Rapid flow reactor AP-20 with a volume of 7L.

[0047] (2) Selection of cells: chicken embryo cell line DF-1, a cell line suitable for the growth of chicken Newcastle disease virus, non-carcinogenic, purchased from the American Type Culture Collection.

[0048] (3) Selection of virus strains: the chicken Newcastle disease virus La sota strain was selected, which can be purchased from the China Veterinary Drug Control Institute.

[0049] (4) Cultivation of different cells: DF-1 cells are cultured in a square bottle, generally passaged at a ratio of 1:3 to 1:5, digested and counted after the cells have grown to a single layer, and used for large-scale cell culture.

[0050] (5) Propagation of cell-adapted virus species: take the cell-adapted Newcastle disease virus virus species, ino...

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Abstract

The invention provides a method for preparing a vaccine by Newcastle disease virus cultured by using a chick embryo continuous cell line and a bioreactor, and belongs to the technical field of animal biological products. The method comprises the following steps: 1) performing passage and culture on a master cell; 2) multiplying a cell-adapted virus seed; 3) culturing the cell for multiplying virus in a suspending manner; 4) culturing and harvesting the virus; 5) preparing a chick Newcastle disease live vaccine; and 6) preparing the chick Newcastle disease live vaccine. The method adopted for preparing vaccines is simple and stable in production process, high in virus content, small in differences among batches, is easy to operate and control the quality, and can remarkably improve the yield and quality of vaccines; the produced chick Newcastle disease vaccines with high safety and high immune efficacy have a complete immune protection effect on the attack of virulent Newcastle disease virus.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a method for preparing a vaccine by cultivating Newcastle disease virus with a chicken embryo passage cell line and a bioreactor. Background technique [0002] Newcastle disease (ND) is a severe viral infectious disease caused by Newcastle disease virus (NDV). It is the most harmful one and is listed as a Class A infectious disease by the International Veterinary Office (OIE). At present, Newcastle disease is still relatively common throughout the country. It restricts the development of my country's poultry industry and the export of poultry products, and also poses potential hazards to food safety and human health. For the control of the disease, in developed countries, hunting of virulent infected groups is the main method, combined with vaccination; while for the majority of developing countries, vaccination is still the main means of contr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/17A61P31/14C12N7/00C12R1/93
Inventor 李阳闫艳丽刘雪王二先徐晓艳盛璐丝
Owner 浙江美保龙生物技术有限公司
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