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Influenza virus splitting vaccine and preparation method thereof

A technology of influenza virus and vaccine, applied in antiviral agents, pharmaceutical formulations, medical preparations containing active ingredients, etc., can solve the problems of influenza virus split vaccines that have not been reported in the literature, achieve high safety in use, and ensure public health Safe and good immune effect

Inactive Publication Date: 2011-05-25
BEIJING MINHAI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Influenza virus split vaccine prepared with CpG ODN adjuvant and aluminum adjuvant has not been reported in the literature at home and abroad

Method used

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  • Influenza virus splitting vaccine and preparation method thereof
  • Influenza virus splitting vaccine and preparation method thereof
  • Influenza virus splitting vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1) Influenza virus proliferated in chicken embryos

[0034] H 1 N 1 、H 3 N 2 , Influenza B virus working seeds were inoculated in 2000 10-day-old healthy chicken embryo allantoic cavities respectively, placed at 34°C, and H 1 N 1 、H 3 N 2 , Type B cultured for 48, 48 and 72 hours respectively;

[0035] 2) Concentration by ultrafiltration

[0036] Place the chicken embryos in a 4°C freezer overnight, and collect H 1 N 1 、H 3 N 2 1. Type B influenza virus allantoic fluid, obtained 23500 milliliters of influenza virus liquid in total, the allantoic fluid was centrifuged at 6000 rpm for 20 minutes, and the supernatant was concentrated to 300 milliliters with an ultrafiltration membrane with a molecular weight cut-off of 300kD;

[0037] 3) centrifugal purification:

[0038] The influenza virus liquid sample concentrated by ultrafiltration was purified by sucrose density gradient centrifugation, monitored by 280nm ultraviolet light, and the virus liquid with viru...

Embodiment 2

[0047] 1) Influenza virus proliferated in chicken embryos

[0048] H 1 N 1 、H 3 N 2 , Influenza B virus working seeds were inoculated in 2000 10-day-old healthy chicken embryo allantoic cavities respectively, placed at 34°C, and H 1 N 1 、H 3 N 2 , Type B cultured for 48, 48 and 72 hours respectively;

[0049] 2) Concentration by ultrafiltration

[0050] Place the chicken embryos in a 4°C freezer overnight, and collect H 1 N 1 、H 3 N 2 1. Influenza virus type B allantoic fluid, 23000 milliliters of influenza virus liquid was obtained, the allantoic fluid was centrifuged at 6000 rpm for 20 minutes, and the supernatant was concentrated to 320 milliliters with an ultrafiltration membrane with a molecular weight cut-off of 300kD;

[0051] 3) Centrifugal purification: The influenza virus liquid concentrated by ultrafiltration is purified by sucrose density gradient centrifugation, monitored by 280nm ultraviolet light, and the virus liquid with virus peaks is collected as...

Embodiment 3

[0058] 1) Influenza virus proliferated in chicken embryos

[0059] H 1 N 1 、H 3 N 2 , Influenza B virus working seeds were inoculated in 2000 10-day-old healthy chicken embryo allantoic cavities respectively, placed at 34°C, and H 1 N 1 、H 3 N 2 , Type B cultured for 48, 48 and 72 hours respectively;

[0060] 2) Concentration by ultrafiltration

[0061] Place the chicken embryos in a 4°C freezer overnight, and collect H 1 N 1 、H 3 N 2 , Type B influenza virus allantoic fluid, obtained 22500 milliliters of influenza virus liquid altogether; The allantoic fluid was centrifuged at 6000rpm for 20 minutes respectively, and the supernatant was concentrated to 340 milliliters with an ultrafiltration membrane with a molecular weight cut-off of 300kD;

[0062] 3) Centrifugal purification: The influenza virus liquid sample concentrated by ultrafiltration was purified by sucrose density gradient centrifugation, monitored by 280nm ultraviolet light, and the virus liquid with v...

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Abstract

The invention provides an influenza virus splitting vaccine and a preparation method thereof. Each dose of influenza virus splitting vaccine contains three influenza hemagglutinins, namely H1N1, H3N2 and type B, a CpG ODN adjuvant and an aluminium adjuvant. The invention also provides a preparation method for the influenza vaccine, which comprises the following steps of: multiplying influenza viruses in chick embryo; performing ultrafitration and concentration; centrifuging and purifying; splitting; performing secondary purification; inactivating; and diluting and packaging. The influenza virus splitting vaccine can reduce the dosage of the influenza virus antigen and production cost of the influenza vaccine, and is suitable for quickly improving influenza vaccine supplying capacity under the threat of global influenza pandemic.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an influenza virus split vaccine containing CpG ODN and an aluminum adjuvant and a preparation method thereof. Background technique [0002] Influenza is a globally prevalent respiratory disease caused by influenza virus. Influenza vaccination is an effective way to prevent influenza. [0003] Traditional influenza virus vaccines include inactivated whole influenza virus vaccines, split influenza virus vaccines and influenza subunit vaccines, all of which are injected intramuscularly without adjuvants. [0004] CpG ODN (oligodeoxynucleotides, ODN) is a single-stranded oligonucleotide containing unmethylated CpG dinucleotide sequence, CpG ODN can be detected by Toll-like receptor 9 (Toll-like receptor 9, TLR- 9) recognized and enhanced immune response. The recombinant hepatitis B vaccine with CpG ODN as an adjuvant of American Dynavax Company has shown in the third phase of clinical...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61K39/39A61P31/16
Inventor 张现臣魏文进黄秋香钟汉斌孟红彦王春雨
Owner BEIJING MINHAI BIOTECH
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