Rabies vaccine virus screening method and preparation method of rabies vaccine

A technology of rabies vaccine and screening method, which is applied in the field of preparation of rabies vaccine and screening of rabies vaccine virus, can solve the problems of long adaptation period, etc., and achieve the effect of increasing yield and shortening adaptation period of subculture

Active Publication Date: 2019-04-23
吉林惠康生物药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The adaptation period of the virus has been shortened by at least 50%, but it still needs 15 generations, and the adaptation period is still long

Method used

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  • Rabies vaccine virus screening method and preparation method of rabies vaccine
  • Rabies vaccine virus screening method and preparation method of rabies vaccine
  • Rabies vaccine virus screening method and preparation method of rabies vaccine

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preparation example Construction

[0040] The invention discloses a preparation method of a rabies virus vaccine, and those skilled in the art can refer to the content of this article and appropriately improve the process parameters to realize it. In particular, it should be pointed out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and the relevant personnel can obviously make changes or appropriate changes and combinations to the method and application described herein without departing from the content, spirit and scope of the present invention to realize and Apply the technology of the present invention.

[0041]The disclosed embodiments are described to enable any person skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those ...

Embodiment 1

[0045] Embodiment 1PM strain rabies virus is respectively subcultured in MRC-5, 2BS cells

[0046] In this example, the PM strain of rabies virus was subcultured in human embryonic lung diploid cells (MRC-5) and human diploid cells (2BS) respectively.

[0047] MRC-5 and 2BS cells were revived in 1.1 and inoculated on T-25cm 2 In a plastic culture flask, after the cells grow into a monolayer, wash with PBS, digest into single cells with 0.25% trypsin, pass down to 25-35 passages at a ratio of 1:2-1:4 for later use.

[0048] 1.2 Inoculate the rabies virus PM strain described in step 1.1 into MRC-5 cells growing into a dense monolayer at an ultra-high MOI of 1000. After infection and adsorption for 0.25-2 hours, discard the virus inoculum and wash with PBS; after washing, add 1 2% fetal bovine serum MEM culture medium containing 1% trehalose, cultured for 2-4 days, then discarded; adding 2% fetal bovine serum MEM culture medium containing 1% trehalose, cultured for 4-6 days to h...

Embodiment 2

[0060] Example 2 CTN strain virus was subcultured in MRC-5 and 2BS cells respectively

[0061] Using the method for passage of virus seeds in Example 1, the CTN strain virus was screened and passaged on MRC-5 and 2BS cells, and the virus titer was detected by direct immunofluorescence method. The results are shown in Table 2.

[0062] Table 2

[0063]

[0064] The results show that when the CTN strain virus is passed to the 6th generation, the virus titer can reach more than 7.5lgFFU / ml, which is 7.0logLD compared with the existing technology when it is passed to the P15th generation 50 / ml method, the present invention greatly shortens the virus adaptation period, and the virus titer is higher.

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Abstract

The invention relates to the technical field of biological medicines, in particular to a rabies vaccine virus screening method and a preparation method of a rabies vaccine. The screening method comprises the step that rabies virus strains are culture in the way that ultrahigh MOI and ultralow MOI are alternatively circulated. Viruses can quickly adapt to cells for six or eight passages, the virustiter is raised to 7.51 gFFU/ml from 4.61 gFFU/ml, the passage adaptation cycle of the virus strains is obviously shortened, the virus yield and the vaccine production efficiency are greatly improved,and the preparation method is suitable for scale production of the rabies vaccine.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a screening method for rabies vaccine virus and a preparation method for rabies vaccine. Background technique [0002] Rabies virus is a neurotropic virus belonging to the genus Lyssavirus in the family Rhabdoviridaes, and is the causative agent of lethal rabies. Almost all warm-blooded animals, including humans, are susceptible to rabies virus. [0003] Rabies, also known as hydrophobia, is an acute zoonotic infectious disease caused by rabies virus infection. Once onset, almost 100% die. WHO recommends that the most effective method is timely vaccination after exposure to rabies virus. It has a history of more than 130 years since the invention of rabies vaccine, from the initial animal nerve tissue vaccine and bird embryo vaccine cell vaccine to today's human diploid cell rabies vaccine (HDCV) and Vero cell rabies vaccine. Today, the most widely used rabies vaccine in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/205A61P31/14C12R1/93
CPCA61K39/12A61P31/14C12N7/00A61K2039/552C12N2760/20134C12N2760/20151
Inventor 袁若森姜春来杨旭张洁琼曾智万铁军何欣魏巍汤鑫原秀娟卢井才张岩
Owner 吉林惠康生物药业有限公司
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