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Method for preparing newcastle disease inactivated vaccine, and product and application thereof

An inactivated vaccine and Newcastle disease technology, which is applied in the direction of medical preparations containing active ingredients, pharmaceutical formulas, virus antigen components, etc., can solve the problems of producing Newcastle disease inactivated vaccines in cell factories, and meet the requirements of immune production and reduce the size of the vaccine. Batch-to-batch differences and the effect of improving product quality

Inactive Publication Date: 2015-06-10
JILIN ZHENGYE BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far there is no report on the use of cell factories to produce inactivated Newcastle disease vaccines

Method used

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  • Method for preparing newcastle disease inactivated vaccine, and product and application thereof
  • Method for preparing newcastle disease inactivated vaccine, and product and application thereof
  • Method for preparing newcastle disease inactivated vaccine, and product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Preparation of Newcastle Disease Inactivated Vaccine by Propagating Newcastle Disease Virus in Cell Factory

[0034] 1. Experimental method

[0035] 1.1 Cell culture

[0036] First, resuscitate the seeded cells DF1 into T75 cell flasks, grow for about 48 hours, and pass passage according to the ratio of 1:4; then transfer the cells in 6 T75 cell flasks full of cells to a 10L spinner bottle for culture; about 48 hours Subculture the cells at a ratio of 1:3; the medium used in this process is DMEM, the serum is fetal bovine serum, and the usage amount is 10%.

[0037] The cells obtained in the spinner bottle were washed twice with PBS, and then 125 ml of 0.25% trypsin-EDTA solution was added for digestion, and the cells in the spinner bottle were harvested.

[0038] After the harvested cells were counted, the DF1 cells were cultured in the Thermo Scientific Nunc cell factory, and the medium used was DMEM medium containing 10% fetal bovine serum. Samples were ...

experiment example 1

[0081] Experimental Example 1 Optimization of Newcastle Disease Virus Proliferation Conditions in Cell Factory

[0082] 1. Experimental method

[0083] 1.1 The effect of cell density on the proliferation of Newcastle disease virus

[0084] At different cell densities (Table 4), the virus was inoculated at MOI=0.2, cultured at 37°C, and the virus was harvested at different time points, and the difference in NDV virus hemagglutination (HA) under different proliferation conditions was compared. The results are shown in Table 4.

[0085] Table 4 Hemagglutination value of virus at different times after inoculation with different cell densities (log2)

[0086]

[0087] As can be seen from Table 4, the cell density was 0.9×10 5 When cells / ml, cultured at 37°C for 48-72h, NDV virus HA reached 8.2log2, better than other treatments. Therefore, the present invention utilizes the cell density of cell factory propagation NDV virus to be determined as 0.9 * 10 5 cells / ml.

[0088] 1...

experiment example 2

[0099] Experimental Example 2 Preparation of Newcastle Disease Inactivated Vaccine with Different Adjuvants and Experiment of Immunity Effect

[0100] 1. Experimental methods and results of adjuvant component screening

[0101] The improved adjuvant A is composed of levamisole, chitosan and complete Freund's adjuvant, 5 mg of levamisole and 8 mg of chitosan in each milliliter of adjuvant, and the balance is complete Freund's adjuvant;

[0102] The improved adjuvant B is composed of levamisole, white oil and saponin, 5 mg of levamisole and 8 mg of saponin in each milliliter of adjuvant, and the balance is ordinary white oil;

[0103] The improved adjuvant C is composed of white oil, chitosan and carbomer (resin), 5 mg of chitosan and 8 mg of carbomer per milliliter of adjuvant, and the balance is ordinary white oil;

[0104] The improved adjuvant D is composed of levamisole, chitosan and white oil. Each milliliter of adjuvant contains 5 mg of levamisole and 8 mg of chitosan, a...

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Abstract

The invention discloses a method for preparing a newcastle disease inactivated vaccine, and a product and application thereof, belonging to the field of preparation and application of newcastle disease inactivated vaccines. The method for preparing the newcastle disease inactivated vaccine disclosed by the invention comprises the following steps of: (1), culturing animal cells by utilizing a cell factory; (2), inoculating newcastle disease virus, and proliferating the virus in the cell factory; and (3), harvesting the virus to prepare the inactivated vaccine. According to the invention, proliferation conditions of the newcastle disease virus in the cell factory including cell density, virus infection multiplicity, TPCK-pancreatin concentration and virus culture time are optimized; the newcastle disease virus cultured by the method disclosed by the invention has high titer, so that immune production requirements can be satisfied; the process is steady, so that the newcastle disease inactivated vaccine can be produced in a large scale; the newcastle disease inactivated vaccine prepared by the method disclosed by the invention is safe to chickens; 100% immunoprotection to attack of the newcastle disease virus can be realized; and thus, the newcastle disease inactivated vaccine can be used for preparing a medicine or a reagent for preventing or treating newcastle diseases.

Description

technical field [0001] The present invention relates to the preparation method of inactivated vaccine, relate in particular to a kind of preparation method of Newcastle disease inactivated vaccine, the present invention also relates to the Newcastle disease inactivated vaccine prepared by said method and application thereof, belong to the preparation and application of Newcastle disease inactivated vaccine application field. Background technique [0002] China has been producing Newcastle disease vaccines from chicken embryos in the past. Because chicken embryos need to consume a lot of chicken embryos to produce Newcastle disease vaccine, the production cycle is long, it is easy to pollute, and it is not easy to control, and each fertilized egg can only be injected with one virus strain at a time, so this production method has low efficiency and is not conducive to large-scale vaccines. large-scale outbreak of Newcastle disease. The prevalence of Newcastle disease in diff...

Claims

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Application Information

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IPC IPC(8): A61K39/17A61K39/39A61P31/14
Inventor 何玉友王国辉田丽华柳志光吴艳丽颜彤王石杨泽斌张微廉维刘斌杨键冯会白杨于志云李海曲红
Owner JILIN ZHENGYE BIOLOGICAL PROD
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