Method for preparing human Japanese encephalitis inactivated vaccine and vaccine

A Japanese encephalitis, inactivated vaccine technology, applied in microorganism-based methods, biochemical equipment and methods, artificial cell constructs, etc. Side effects, low safety and other problems, to achieve the effect of reducing pollution, improving vaccine safety, and reducing pollution

Pending Publication Date: 2022-04-05
LIAONING CHENGDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Among them, the Vero cell inactivated Japanese encephalitis vaccine effectively reduces the incidence of Japanese encephalitis after human infection with JEV; however, although the inactivated Japanese encephalitis vaccine has undergone certain treatments, it still contains a small amount of allogeneic Proteins can stim

Method used

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  • Method for preparing human Japanese encephalitis inactivated vaccine and vaccine
  • Method for preparing human Japanese encephalitis inactivated vaccine and vaccine
  • Method for preparing human Japanese encephalitis inactivated vaccine and vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Serum-free adaptation of Vero cells

[0059] 1. Acclimatization of Vero cells

[0060] In this example, Vero cells derived from ATCC were acclimatized without serum using a mixed medium, and the specific operation steps are as follows:

[0061] (1) Recovery of Vero cells;

[0062] (2) domesticate Vero cells in the mixed culture medium;

[0063] Step 1: Culture the revived Vero cells in serum-containing medium. After the cells grow to a monolayer, wash the cells with DPBS, and then digest the cells with recombinant trypsin for passage;

[0064] Step 2: Culture the revived Vero cells in the first mixed solution. After the cells grow to a monolayer, wash the cells with DPBS, and then digest the cells with recombinant trypsin for passage;

[0065] Step 3: Culture the revived Vero cells in the second mixed solution. After the cells grow to a monolayer, wash the cells with DPBS, and then digest the cells with recombinant trypsin for passage;

[0066] Step 4: Cu...

Embodiment 2

[0083] Embodiment 2: the propagation of Japanese encephalitis virus in serum-free culture system

[0084] In the production of the Japanese encephalitis inactivated vaccine, the step of inoculating JEV on Vero cells to proliferate is included. In this example, a method for the propagation of Japanese encephalitis virus in a serum-free culture system is provided. In this method, JEV is inoculated onto Vero cells adapted to serum-free medium cultured in Example 1 , the specific operation steps are as follows:

[0085] Step 1, cell proliferation: culture the Vero cells acclimated in Example 1 in a serum-free medium, and discard the supernatant after the cells grow to a monolayer;

[0086] Step 2, virus inoculation: according to MOI=0.001-0.1, insert JEV and adsorb for 1 hour;

[0087] Step 3. Virus cultivation: add maintenance solution after virus inoculation, place at 33°C, 5% CO 2 Cultivate in an incubator;

[0088] Step 4. Virus harvest: change the medium after 1 day of cu...

Embodiment 3

[0095] Embodiment 3: Japanese encephalitis inactivated vaccine for humans

[0096] 1. Preparation of Inactivated Japanese Encephalitis Vaccine for Human

[0097] In this example, the Vero seed cells adapted to the serum-free medium obtained using the method described in Example 1 were added to the bioreactor, and the perfusion culture was carried out together with the microcarriers in the serum-free medium, and then the microcarriers were settled. Carriers and cells, replace the medium with maintenance solution, inoculate JEV to continue perfusion culture, collect the virus solution, and then concentrate, filter, inactivate, purify and freeze-dry to obtain the inactivated Japanese encephalitis vaccine for human use , the specific operation steps are as follows:

[0098]Step 1. Expansion culture of Vero cells: the Vero seed cells adapted to the serum-free medium obtained by acclimating using the method described in Example 1 were divided into 1.0×10 6 The inoculation density ...

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Abstract

The invention discloses a method for preparing a human encephalitis B inactivated vaccine and the vaccine, and belongs to the field of vaccine manufacturing, the method for preparing the human encephalitis B inactivated vaccine sequentially comprises the following steps: S1, serum-free adaptive domestication of Vero cells on a culture flask; s2, performing serum-free multiplication culture on the Vero cells on the bioreactor; s3, inoculating JEV on the Vero cells in the bioreactor; s4, harvesting, inactivating, concentrating and purifying viruses; s5, carrying out freeze-drying treatment on the vaccine stock solution; the Japanese encephalitis inactivated vaccine has the effects of improving the safety of the Japanese encephalitis inactivated vaccine for people and reducing the side effects of the Japanese encephalitis inactivated vaccine for people.

Description

technical field [0001] The application relates to the field of vaccine manufacture, in particular to a method for preparing an inactivated Japanese encephalitis vaccine for humans and the vaccine. Background technique [0002] Japanese encephalitis, referred to as Japanese encephalitis, is a zoonotic mosquito-borne infectious disease caused by Japanese encephalitis virus (JEV). And Japanese encephalitis is extremely harmful, and is listed as one of the key infectious diseases by the World Health Organization. [0003] JEV infection initially manifests as atypical high fever, and the course of the disease usually lasts for 2-3 weeks. The main clinical manifestations are high fever, headache, coma, tremor, movement disorder, and nervous system deficits. Focal onset; immune-mediated transverse myelitis, acute disseminated encephalomyelitis, and NMDA encephalitis may follow infection. [0004] At present, there is still no effective drug to treat Japanese encephalitis infectio...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61P31/14C12N5/071C12N7/00C12R1/93
Inventor 李庆岸杨莹莹王立刚陈佳丽杨健由迪
Owner LIAONING CHENGDA BIOTECH
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