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Combined vaccine, and preparation method and application thereof

A combination vaccine and inactivation technology, applied in the biological field, can solve the problems of inability to use attenuated vaccines in combination, differences in mechanism of action, cumbersome vaccination procedures, etc., and achieve the effect of reducing the number of vaccinations

Active Publication Date: 2020-02-04
ANHUI ZHIFEI LONGCOM BIOPHARM CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, in order to prevent Japanese encephalitis and chickenpox, two different vaccines need to be inoculated separately, that is, Japanese encephalitis vaccine or chickenpox vaccine, and the vaccination procedure is cumbersome.
The combined use of the existing Japanese encephalitis live attenuated vaccine and varicella live attenuated vaccine has a greater risk, because the attenuated live vaccine always has risks such as recombination with wild strains, self-mutation, and shedding of live virus to nature. For example: VZV attenuated strains have infected normal people
Moreover, the immunization procedures of the two existing attenuated vaccines are different, and the mechanism of action is different, so that the two attenuated vaccines cannot be used in combination

Method used

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  • Combined vaccine, and preparation method and application thereof
  • Combined vaccine, and preparation method and application thereof
  • Combined vaccine, and preparation method and application thereof

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preparation example Construction

[0060] The present invention also includes a preparation method of the combined vaccine, the preparation method comprising the following steps:

[0061] Preparation of inactivated varicella-zoster virus purified liquid;

[0062] Prepare the purified solution of inactivated Japanese encephalitis virus;

[0063] Mix the inactivated Japanese encephalitis virus purified liquid and the inactivated varicella-zoster virus purified liquid,

[0064] Sterilize and freeze-dry the mixed purified virus liquid to obtain the combined vaccine.

[0065] The steps for preparing the purified solution of inactivated varicella-zoster virus are as follows:

[0066] Human diploid cells are subcultured;

[0067] Inoculating the varicella-zoster virus onto the subcultured human diploid cells, and culturing the virus to obtain a varicella-zoster virus harvest liquid;

[0068] Concentrate the varicella-zoster virus harvest liquid by ultrafiltration until the total protein content in the concentrated...

Embodiment 1

[0094] Embodiment 1 The preparation method of inactivated varicella-zoster virus purified liquid

[0095] Experimental materials and sources:

[0096] Human diploid ZFB-3 cells;

[0097] Varicella zoster virus, clinical collection;

[0098] 0.2%~0.5% human serum albumin, purchased from Tonglu Biopharmaceutical;

[0099] MEM medium, purchased from Thermo company;

[0100] PBS, homemade;

[0101] Ultrafiltration membrane bag, purchased from Millipore;

[0102] β-propiolactone, purchased from Serva company;

[0103] Preparation:

[0104] 1. Passage ZFB-3 cells to 20 generations in cell culture flasks and cell factories, wherein ZFB-3 cells are host cells;

[0105] 2. When the host cell is amplified to a fusion degree of more than 90%, the varicella-zoster virus is inoculated into the host cell for infection, and the M.O.I. is 0.001;

[0106] 3. Add MEM medium containing 0.2% to 0.5% human serum albumin, and place it in a cell incubator at 34°C for cultivation;

[0107] 4...

Embodiment 2

[0113] The adaptation of embodiment 2 Japanese encephalitis virus on ZFB-3 cell

[0114] Experimental materials and sources:

[0115] Human diploid ZFB-3 cells;

[0116] Japanese encephalitis virus (JEV) P3 strain was purchased from China National Institutes for Food and Drug Control

[0117] Fetal bovine serum was purchased from GIBCO and Hangzhou Sijiqing;

[0118] MEM medium, purchased from Thermo company;

[0119] PBS, Homemade.

[0120] experiment procedure:

[0121] The mouse brain-derived Japanese encephalitis virus P3 strain was adapted to MRC-5 cells and ZFB-3 cells respectively;

[0122] Using the method of multiple screening of virus plaques, the P3 virus strains should not be less than 10 -3 After doubling dilution, they were seeded in 96-well plates of monolayer cells, and MRC-5 cells and ZFB-3 cells were respectively set to correspond to 96-well plates. Adaptation on MRC-5 cells and ZFB-3 cells was carried out simultaneously under the same conditions.

[...

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Abstract

The invention relates to a combined vaccine, and a preparation method and application thereof. The combined vaccine comprises an inactivated japanese encephalitis virus and an inactivated varicella zoster virus; the ratio of the total protein content of the inactivated japanese encephalitis virus in the combined vaccine to the total protein content of the inactivated varicella zoster virus in thecombined vaccine is (0.25-4):1; the preparation method of the combined vaccine comprises the steps that an inactivated japanese encephalitis virus purification solution and an inactivated varicella zoster virus purification solution are mixed to obtain a mixed virus purification solution; and the inactivated japanese encephalitis virus and the inactivated varicella zoster virus are prepared by adopting the same human diploid cells as a virus culture medium to realize mixture of the inactivated japanese encephalitis virus and the inactivated varicella zoster virus to obtain the combined vaccine. The prepared combined vaccine is safe and effective, and the number of vaccinations is effectively reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a combined vaccine and its preparation method and application. Background technique [0002] At present, there are mainly two types of Japanese encephalitis vaccines: live attenuated Japanese encephalitis vaccine (hamster kidney cells) and inactivated Japanese encephalitis vaccine (Vero cells). At present, there are three types of JE vaccines used on a large scale: (1) Nakayama strain or Beijing strain purified mouse brain inactivated vaccine, mainly produced in several Asian countries such as Japan; (2) Beijing P3 strain JE inactivated by Vero cells Vaccine; (3) SA14-14-2 strain live attenuated vaccine cultured by primary hamster kidney cells. There are live attenuated Japanese encephalitis vaccines (hamster kidney cells) and inactivated Japanese encephalitis vaccines (Vero cells) on the market in my country. [0003] Because the existing Japanese encephalitis live attenuated vacc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/295A61K39/12A61K39/25A61P31/14A61P31/22C12N7/00C12N7/06C12R1/93
CPCA61K39/12A61P31/14A61P31/22C12N7/00A61K2039/70A61K2039/5252C12N2710/16734C12N2770/24134C12N2710/16751C12N2710/16763C12N2770/24151C12N2770/24163Y02A50/30
Inventor 蒋仁生谭小东杨世龙沈慧莉张艳飞
Owner ANHUI ZHIFEI LONGCOM BIOPHARM CO LTD
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