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Human varicella virus inactivated vaccine and preparation method thereof

A technology for varicella virus and inactivated vaccines, applied in biochemical equipment and methods, viruses, antiviral agents, etc., can solve problems affecting the quality of vaccines, and achieve economic benefits, good immunogenicity, and thorough inactivation effects Effect

Active Publication Date: 2020-11-27
ANHUI ZHIFEI LONGCOM BIOPHARM CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Poor control of dose uniformity in the irradiation process will also affect the quality of the vaccine

Method used

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  • Human varicella virus inactivated vaccine and preparation method thereof
  • Human varicella virus inactivated vaccine and preparation method thereof
  • Human varicella virus inactivated vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1 preparation method of human varicella virus inactivated vaccine

[0037] (1) MRC-5 cells are subcultured to 28 generations in cell culture flasks and cell factories;

[0038](2) When the cells are amplified to a confluence of more than 90%, the virus is inoculated into the cells for infection, and the M.O.I. is 0.01-0.1;

[0039] (3) Add MEM medium containing 2% to 5% fetal bovine serum, and place in a cell incubator at 34 to 37°C for cultivation;

[0040] (4) observe the degree of cell lesion every day, and when the CPE of the cells reaches more than 50%, discard the culture medium and wash the cells with PBS;

[0041] (5) Add 0.03% EDTA solution to elute the infected cells, collect the cell suspension in a centrifuge tube, centrifuge at 4000rpm for 10 minutes at 4°C, remove the supernatant, add a stabilizer to combine, mix well, and take a sample from a single bottle. Bacteria detection, frozen at -80°C;

[0042] (6) Place the sample to be tested for s...

Embodiment 2

[0049] The selection of embodiment 2 virus concentrated liquid protein concentration

[0050] (1) Prepare 5 batches of inactivated virus liquids according to steps (1)-(10) of the preparation method of human varicella virus inactivated vaccine;

[0051] (2) The measured concentration of the virus concentrate and the infectivity after inactivation are shown in the following table:

[0052]

[0053] According to the results in the above table, we chose to inactivate the virus concentrate with a protein content of 35-95 mg / ml.

Embodiment 3

[0054] Embodiment 3 plaque method measures the infectivity of varicella virus inactivated vaccine

[0055] Spread MRC-5 cells on a 6-well plate, inoculate 3*10 per well 5 cells, 5% CO 2 After incubating in an incubator at 34-37°C for 3-4 days, the titer of varicella virus can be measured after the cells grow into a monolayer. Aspirate the medium, dilute the inactivated varicella virus vaccine to an appropriate concentration to infect the cells, and in 5% CO 2 Adsorb in an incubator at 34-37°C for 1-2 hours, then add 3 mL of virus culture solution to each well, and store in 5% CO 2 Cultivate in an incubator at 34-37°C. Staining was carried out after 7-10 days of culture. Take the average value of the counts, multiply it by the dilution factor and the inoculation volume correction factor, and obtain the titer of the inactivated varicella virus vaccine, which is expressed in lg PFU / mL. The titer results of the inactivated varicella virus inactivated vaccine are all empty spo...

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Abstract

The invention relates to the field of biotechnology, and particularly provides a varicella virus inactivated vaccine used for people and a preparation method thereof. The preparation method comprises the following steps: mixing beta-propiolactone and varicella virus concentrated solution at a volume ratio of (1:2000) to (1:8000), carrying out inactivation for 12-36h at a temperature of 3-5 DEG C; then, at the temperature of 34-37 DEG C, carrying out hydrolysis for 1.5-3h to obtain inactivated virus liquid; carrying out purification, split charging and freeze-drying on the inactivated virus liquid to obtain the inactivated vaccine. The varicella virus inactivated vaccine prepared with the method is safe and effective, is slight in vaccination reaction and is more suitable for crowds who can not use attenuated live vaccines and have poor immunologic functions.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a human varicella virus inactivated vaccine and a preparation method thereof. Background technique [0002] Varicella-zoster virus (VZV) is the causative agent of both chickenpox and herpes zoster. VZV belongs to the Herpesviridae family and is a double-stranded DNA virus. [0003] VZV is a highly contagious herpes virus that causes both varicella and herpes zoster (HZ); the former is usually seen in childhood and young adults, while the latter is a result of reactivation and replication of latent VZV lead to. After initial infection with VZV, the virus can remain dormant in sensory ganglia, and such reactivation of VZV is associated with decreased immune function, mostly in elderly or immunocompromised populations (Weinberg et al., Journal of Infectious Diseases (2009 ) 200:1068-77). Some patients have pain associated with HZ that persists long after the HZ rash has healed. [...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/25A61P31/22A61L2/18A61L101/36
CPCA61K39/12A61K2039/5252A61K2039/55A61L2/0088A61L2202/21C12N2710/16734
Inventor 蒋仁生谭小东张艳飞沈慧莉杨世龙
Owner ANHUI ZHIFEI LONGCOM BIOPHARM CO LTD
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