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Multivalent immunogenic composition containing enterovirus antigens

A technology of immunogenicity and viral antigen, applied in the direction of viral antigen components, antiviral agents, drug combinations, etc., can solve time-consuming and labor-intensive problems

Active Publication Date: 2015-06-17
SINOVAC BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both the hand, foot and mouth disease vaccine under development and the polio vaccine on the market are multi-dose immunization, and the recipients need to be vaccinated multiple times, which is time-consuming and labor-intensive

Method used

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  • Multivalent immunogenic composition containing enterovirus antigens
  • Multivalent immunogenic composition containing enterovirus antigens
  • Multivalent immunogenic composition containing enterovirus antigens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] The cultivation of embodiment 1EV71 virus

[0069] The EV71 virus strain (preservation number is CGMCC No.3544) was adapted to grow in 10-layer cell factory Vero cells and continuously passaged, using MEM medium plus 10% (v / v) calf serum, and cultured at 37±1°C After culturing in the box for 5-7 days, change the MEM medium and add 2% (v / v) calf serum, inoculate EV71 virus according to the MOI of 0.001, culture at 33±1°C for 15 days, observe the cytopathic changes every day, when the cytopathic changes reach 50 % or more, the virus liquid was harvested, and after clarification and filtration, the virus titer value was detected by the CCID50 method. The EV71 virus harvest liquid can be in the range of 6-8LgCCID50 / ml.

Embodiment 2

[0070] Inactivation and purification of embodiment 2EV71 virus

[0071] Add formaldehyde solution (1:1000-1:10000) (v / v) to the EV71 virus harvest solution described in Example 1, so that the final concentration of free formaldehyde is 100-200 μg / ml, and inactivate at 36.5°C±1°C On day 13, a virus inactivation solution was prepared. Centrifuge the inactivation solution at 30,000rpm in a sucrose density gradient for 6-12 hours, collect the zone where the virus is located, and desugar through membrane bag ultrafiltration. After molecular sieve chromatography, equilibrate with 0.01M PBS (pH7.0) buffer, then continue to elute with 0.01M PBS (pH7.0), collect virus peaks, and finally obtain EV71 virus antigen stock solution after sterile filtration.

Embodiment 3

[0072] The cultivation of embodiment 3CA16 virus

[0073] The CA16 virus strain (preservation number is CGMCC No.5371) was adapted to grow in 10-layer cell factory Vero cells and continuously passaged, using MEM medium plus 10% calf serum, and cultured in a 37±1°C incubator for 5- After 7 days, change the MEM medium and add 2% calf serum, inoculate CA16 virus according to the MOI of 0.001, and continue to culture in the incubator at 37±1°C, observe the cytopathic changes every day, and harvest the virus liquid when the cytopathic changes reach more than 50% , After clarification and filtration, the virus titer value was detected by the CCID50 method. CA16 virus harvest liquid can be in the range of 6-8LgCCID50 / ml.

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Abstract

The invention provides a multivalent immunogenic composition containing enterovirus antigens. The composition comprises inactivated EV71 antigens and / or inactivated CA16 antigens, and inactivated polio antigens. The composition can further comprise antigens selected from hepatitis A antigens, hepatitis B antigens, acellular pertussis antigens, tetanus toxoid, diphtheria toxoid, Haemophilus influenzae type b capsular polysaccharide, and meningococcal polysaccharide antigens, as well as physiologically acceptable carriers combined with bacterial polysaccharide antigens. The invention also provides a preparation method of the composition. The composition can prevent invasion of a plurality of pathogens simultaneously without interference among the antigens, and the immunogenicity is no less than that of individually activated antigens. With the composition, vaccination processes are significantly simplified, and the vaccination efficiency is improved with reduced costs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a multivalent immunogenic composition containing enterovirus antigens. Background technique [0002] (1) HFMD and HFMD pathogens [0003] Hand, foot and mouth disease is a common, mild, but frequent and highly contagious disease. Hand, foot and mouth disease can take place throughout the year, more common in summer and autumn. Most patients have mild symptoms, mainly fever, rashes and herpes on the hands, feet and mouth. In a small number of cases (especially those younger than 3 years old), encephalitis, encephalomyelitis, meningitis, pulmonary edema, and circulatory failure may occur. Individual children with severe disease develop rapidly and easily lead to death. HFMD can be caused by a variety of enteroviruses, among which Enterovirus EV71 and Coxsackievirus A16 (CA16) are the main epidemic strains. [0004] Human enterovirus type 71 (EV71) belongs to the Picornaviridae fam...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/13A61K39/125A61K39/295A61K39/29A61K39/10A61K39/05A61K39/08A61K39/02A61K39/102A61K39/095A61P31/14A61P1/00
CPCY02A50/30
Inventor 王一丁姜德玉赵崇伯戈小琴韩星董珊珊高强尹卫东
Owner SINOVAC BIOTECH
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