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Combined EV71 (enterovirus 71)-HA (hepatitis A) vaccine

An enterovirus and combined vaccine technology, applied in the field of medical biology, to achieve the effect of reducing the number of vaccinations, reducing the production cost, and having a good conformation

Active Publication Date: 2013-05-08
SINOVAC BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

HAV does not cause chronic infection, and currently there is no specific antiviral treatment

Method used

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  • Combined EV71 (enterovirus 71)-HA (hepatitis A) vaccine
  • Combined EV71 (enterovirus 71)-HA (hepatitis A) vaccine
  • Combined EV71 (enterovirus 71)-HA (hepatitis A) vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Preparation of inactivated EV71 stock solution

[0024] Cultivate the Vero cells to be inoculated until they grow into a dense monolayer, then digest the cells, subculture at 1:8 and inoculate the virus until they grow into a monolayer, and inoculate the virus with the EV71 inactivated vaccine virus seeds at a ratio of 1:10 2 CCID 50 / ml inoculated in the well-grown Vero cells, cultured at 33±1°C for 15 days, and harvested the cultured cell virus supernatant according to the cell pathological changes to obtain EV71 virus harvest liquid. The harvest liquid is inactivated by formaldehyde with a final concentration of not more than 250 μg / ml for not less than 6 days, and then purified by ultrafiltration and column chromatography, and the combined chromatographic purified liquid is concentrated by ultrafiltration and then sterilized and filtered to obtain inactivated EV71 stock solution.

Embodiment 2

[0025] Embodiment 2: Preparation of inactivated HAV stock solution

[0026] 2BS cells were cultured in Eagle's MEM medium containing newborn bovine serum. After the cells had grown well, they were digested and subcultured by dilution, and then inoculated with HAV of TZ84 strain, YN5 strain, HM175 strain and Lv8 strain at a MOI of 0.05-1.0. During the period, the maintenance medium was changed according to the growth of the cells, and the maintenance medium was Eagle's MEM medium containing newborn bovine serum. After culturing to the peak of virus proliferation, trypsin or other appropriate methods are used to digest cells containing HAV, and the virus harvest liquid is collected by centrifugation or filtration. The virus is concentrated by ultrafiltration, and then purified by chromatography; after sterilization and filtration, it is inactivated with formaldehyde with a final concentration of no more than 250 μg / ml at 37±1°C for 12 days, and the inactivated virus liquid is in...

Embodiment 3

[0027] Embodiment 3: Preparation of EV71 and HAV combined vaccine

[0028] Adsorption method 1: After the diluted inactivated HAV and the diluted inactivated EV71 are adsorbed with aluminum adjuvant respectively, the adsorption products of the two inactivated viruses are mixed to prepare a combined vaccine of EV71 and HAV.

[0029] Adsorption method 2: The diluted inactivated HAV is first adsorbed with an aluminum adjuvant, and then diluted inactivated EV71 is added for adsorption to prepare a combined vaccine of EV71 and HAV.

[0030] Adsorption method 3: The diluted and inactivated EV71 is first adsorbed with aluminum adjuvant, and then diluted and inactivated HAV is added for adsorption to make a combined vaccine of EV71 and HAV.

[0031] Adsorption method 4: Mix the diluted inactivated HAV and the diluted inactivated EV71 first, and then add the mixed solution to the aluminum adjuvant for adsorption to make a combined vaccine of EV71 and HAV. The content of EV71 antigen i...

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Abstract

The present invention discloses a combined EV71 (enterovirus 71)-HA (hepatitis A) vaccine which comprises inactivated enterovirus 71 viruses, inactivated hepatitis A viruses, and a aluminium adjuvant. The invention also provides a method for preparing the combined EV71-HA vaccine; and in the method, the protective agents such as gelatins and the like are not required to be added, the pH adjustment is not required to be performed, and the absorption effect and stability of the prepared vaccine are good. After the vaccine is vaccinated, the hepatitis A virus has an adjuvant effect on EV71 (enterovirus 71), therefore, the immunogenicity of the EV71 (enterovirus 71) is enhanced, and the immunity effect of the combined EV71-HA vaccine is equal to or greater than the immunity effect of a univalent vaccine. The vaccine disclosed by the invention has EV71-HA double immunities and protection performances; when the vaccine disclosed by the invention is vaccinated, the inoculating needle time can be reduced, and the immunization process can be simplified; and meanwhile, the diseases of people and animals arising from enterovirus 71 viruses and hepatitis A viruses can be effectively prevented.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and relates to a combined vaccine and a preparation method thereof, in particular to a combined vaccine of enterovirus 71 and hepatitis A and a preparation method thereof. Background technique [0002] In recent years, the incidence of hand, foot and mouth disease has become more and more frequent, and the number of cases and deaths has increased year by year, which seriously threatens the health of children and infants in my country and causes a huge economic burden to the society. In May 2008, my country included HFMD in the management of Class C infectious diseases. Enterovirus 71, as the most important pathogen of HFMD, is often associated with large-scale outbreaks, severe cases and death cases of HFMD. Hepatitis A is a major highly contagious intestinal infectious disease with a high incidence and a wide range of prevalence. It is a Class B infectious disease under legal management in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/295A61P31/14A61K39/125A61K39/29
CPCY02A50/30
Inventor 胡伟蔡芳薛晨宝高强尹卫东
Owner SINOVAC BIOTECH
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