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Virus split deactivation method for human influenza virus split vaccine

An influenza virus inactivation technology, applied in antiviral agents, pharmaceutical formulations, inactivation/attenuation, etc., can solve the problems of poor uniformity and stability of inactivation, poor virus lysis effect, and many inactivation steps , to achieve the effect of shortening the inactivation time, improving the cracking effect, uniform and thorough cracking

Active Publication Date: 2013-10-30
长春生物制品研究所有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The invention provides a virus lysis and inactivation method for influenza virus cleavage vaccines for human use, so as to solve the problems of poor virus lysis effect, long inactivation time, low inactivation efficiency and large amount of inactivation agent existing in the current traditional pre-inactivation process , There are many inactivation steps, the risk of bacterial contamination is increased, the plant, facilities and energy consumption required for inactivation are increased, and the uniformity and stability of inactivation are poor

Method used

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  • Virus split deactivation method for human influenza virus split vaccine
  • Virus split deactivation method for human influenza virus split vaccine
  • Virus split deactivation method for human influenza virus split vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Production of monovalent stock solution of H1N1 influenza virus

[0047] 1) Chicken embryos for production

[0048] The chicken embryos used for subculture and preparation of virus seeds should come from SPF chickens; the chicken embryos used for vaccine production should come from healthy chickens raised in closed houses, and 9-11 days old chickens with no deformities, clear blood vessels, and activities should be selected. Embryo.

[0049] 2) Virus inoculation and cultivation

[0050] Use PBS (0.01 mol / L, pH 7.2) to dilute the working seed batch virus seeds, and the H1N1 virus seeds are diluted to 4.0LgEID50 / ml. Inoculate the allantoic cavity of chicken embryos, 0.2ml per embryo. Incubate at 33-35°C for 44-48 hours. If the working seeds are not used up after being thawed, they should not be frozen for further use.

[0051] 3) Virus Harvest

[0052] Screen the live chicken embryos, place the embryos at 2-8°C for 12-18 hours, harvest the allantoic fluid ...

Embodiment 2

[0083] Example 2 Production of monovalent stock solution of H3N2 influenza virus

[0084] 1) Chicken embryos for production

[0085] The chicken embryos used for subculture and preparation of virus seeds should come from SPF chickens; the chicken embryos used for vaccine production should come from healthy chickens raised in closed houses, and 9-11 days old chickens with no deformities, clear blood vessels, and activities should be selected. Embryo.

[0086] 2) Virus inoculation and cultivation

[0087] Use PBS (0.01 mol / L, pH 7.2) to dilute the batch virus seeds of the working seeds, dilute the H3N2 type virus seeds to 4.0LgEID50 / ml, inoculate the allantoic cavity of chicken embryos, inoculate 0.2ml per embryo, and culture at 33-35°C for 44- For 48 hours, if the working seeds and poisonous seeds are melted, if they are not used up at one time, they must not be frozen for further use.

[0088] 3) Virus Harvest

[0089] Screen the live chicken embryos, place the embryos at ...

Embodiment 3

[0120] Example 3 Production of Influenza B Monovalent Stock Solution

[0121] 1) Chicken embryos for production

[0122] The chicken embryos used for subculture and preparation of virus seeds should come from SPF chickens; the chicken embryos used for vaccine production should come from healthy chickens raised in closed houses, and 9-11 days old chickens with no deformities, clear blood vessels, and activities should be selected. Embryo.

[0123] 2) Virus inoculation and cultivation

[0124] Use PBS (0.01 mol / L, pH7.2) to dilute the working seed batch virus, dilute the type B virus to a virus amount of 3.0 LgEID50 / ml, inoculate the allantoic cavity of chicken embryos, inoculate 0.2ml per embryo, and place at 33- Incubate at 35°C for 66-72 hours. If the working seeds have been thawed and are not used up once, they must not be frozen for further use.

[0125] 3) Virus Harvest

[0126] Screen the live chicken embryos, place the embryos at 2-8°C for 12-18 hours, harvest the al...

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Abstract

The invention relates to a virus split deactivation method for a human influenza virus split vaccine, and belongs to the field of biological pharmacy. A method of firstly purifying and cracking H1N1 type, H3N2 type or B type monovalent influenza totivirus after gradient density centrifugal purification and then deactivating is adopted, so that the virus cracking effect can be improved, the deactivation time can be shortened, the deactivation efficiency can be increased, the using amount of a deactivator can be obviously reduced, deactivation operation steps can be simplified, the contamination risk can be reduced, factory buildings, facilities and energy consumption required for deactivation can be reduced, and the uniformity and the stability of deactivation can be increased.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, in particular to a method for cracking and inactivating influenza virus monovalent stock solution in the production of human influenza virus split vaccine. Background technique [0002] Influenza (flu for short) is an acute respiratory infectious disease caused by influenza virus (Influenza virus). In recent years, the continuous outbreak of avian influenza (Avian influenza) and the outbreak of new type A H1N1 influenza in 2009 have further increased the pressure on health and epidemic prevention in various countries, causing people panic. Influenza is the first infectious disease to achieve global surveillance, but there is no specific drug for its treatment. Vaccination is an effective measure to prevent influenza epidemics. Influenza virus is an enveloped virus. The inside of the virus particle is a nucleocapsid, which is composed of nucleoprotein, polymerase, and viral genomic RNA; the env...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/145A61P31/16C12N7/06
Inventor 张雪梅王亚军赵大鹏李晓波张赐强徐文顾建阳范宇红匡科伟毕业任琦刘荫郭占龙赵志芳郭雪刘旭光孙丽娟王春鹏张玉辉宋泽民韩俊海谷丽娟吴业宏
Owner 长春生物制品研究所有限责任公司
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