Combined vaccine for preventing hand-foot-mouth disease

A combined vaccine and hand, foot and mouth disease technology, applied in vaccines, multivalent vaccines, viruses, etc., can solve the problems of unknown immune interference, reduce vaccine single-component immunogenicity, affect immune effect, etc., and achieve good process consistency , Improving vaccination efficiency and simplifying vaccination procedures

Active Publication Date: 2016-09-28
SINOVAC BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant report on the development of a combined vaccine of EV71, CA16 and CA10. First, a large-scale outbreak of hand, foot and mouth disease caused by EV71, CA16 and CA10 virus has occurred in a relatively short period of time. Secondly, hand, foot and mouth disease caused by virus There is a certain regional distribution. For developed countries with a relatively low incidence, there are few institutions investing in research and development. Third, the immune interference between different viral antigens is unknown, and this interference may directly reduce the efficacy of a single-component vaccine. Immunogenicity, affecting immune effect

Method used

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  • Combined vaccine for preventing hand-foot-mouth disease
  • Combined vaccine for preventing hand-foot-mouth disease
  • Combined vaccine for preventing hand-foot-mouth disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The preparation of embodiment 1 virus liquid

[0047] 1. Cell culture and recover SLF-1 cells, use MEM solution containing 20% ​​bovine serum, culture at 37±1°C, replace the medium after adherence, and use MEM solution containing 10% bovine serum. Wash the monolayer, digest with 0.25% trypsin, and divide into culture flasks or cell factories at a ratio of 1:4, transfer to 40-layer cell factories, grow to a monolayer, and prepare for inoculation of viruses.

[0048] 2. Virus culture

[0049] The cells were inoculated with EV71 virus at an MOI of 0.0001, cultured at 36±1°C with a culture medium containing 2% bovine serum, and the virus was harvested after 8 days.

[0050] The CA16 virus was inoculated into the cells at an MOI of 0.001, cultured at 35±1°C with a culture solution containing 1% bovine serum, and the virus was harvested after 10 days.

[0051] The CA10 virus was inoculated into the cells at an MOI of 0.001, cultured at 35±1°C in culture medium without bovin...

Embodiment 2

[0052] The inactivation of embodiment 2 virus

[0053] (1) Clarification: Filter the virus liquid through a filter element with a continuous two-stage pore size of 3-0.8 μm and 0.65-0.22 μm or centrifuge (including continuous flow centrifugation) with a rotation speed of 3000 rpm and a centrifugation time of 0.5 hours to obtain a clarified virus liquid;

[0054] (2) Inactivation Divide EV71, CA16, and CA10 virus clarified liquid into 9 groups respectively, add different volumes of 1:200 formalin solution to each group, so that the theoretical final concentration is 200, 100, 67 μg / ml, each Concentrations of 3 groups of parallel samples were inactivated at 37±1°C. See Table 1 for grouping information. Sampling was carried out at 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, and 24 hours of inactivation, and immediately after the sampling was completed, formaldehyde was neutralized with sodium bisulfite solution, and virus titration was carried out. degree measurement. The virus ...

Embodiment 3

[0064] The purification of embodiment 3 virus

[0065] (1) Ultrafiltration concentration of virus liquid Select ultrafiltration membrane packs with molecular weight cut-offs of 100kD, 300kD, and 500kD three pore sizes, respectively carry out ultrafiltration concentration of EV71 (or CA16 or CA10) with a batch of inactivation liquid, and fix the ultrafiltration pressure , concentrate the same multiple (100 times), fix the number of times of diafiltration (8 times of diafiltration), compare the ultrafiltration time and the antigen recovery rate of three membrane packs, the removal rate of impurity protein and the activity ratio of the product before and after ultrafiltration concentration (antigen content / protein content), and detect the antigen content of each filtrate. Determine the pore size of the ultrafiltration membrane package according to the test results. The results are shown in Table 4.

[0066] Table 4 Comparison results of ultrafiltration effects with different m...

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Abstract

The invention provides a combined vaccine for preventing a hand-foot-mouth disease. The combined vaccine contains an inactivated EV71 (Enterovirus 71) antigen, an inactivated CA16 (Coxasckievirus A16) antigen and an inactivated CA10 (Coxasckievirus A10) antigen. The hand-foot-mouth disease preventing combined vaccine with good immunogenicity, high safety and high stability is prepared by culturing main epidemic virus strains, namely an EV71 type, a CA16 type and a CA10 type, which cause the hand-foot-mouth disease, performing ultrafiltration concentration, performing sucrose density gradient centrifugation, performing ultrafiltration desugarization and performing sterilization filtration to obtain a purified virus solution, and further preparing the virus solution into the combined vaccine. The combined vaccine provided by the invention can prevent infection of a variety of enteroviruses to a human body at the same time, and a phenomenon of mutual interference among the antigens does not exist and the corresponding immunogenicity is not lower than that excited by a single antigen; after use of the combined vaccine provided by the invention, a vaccinating procedure can be significantly simplified, the vaccinating efficiency is improved and the cost is reduced.

Description

technical field [0001] The invention relates to the technical field of preparation of biological products, in particular to a combined vaccine containing enterovirus antigens for preventing hand, foot and mouth disease. Background technique [0002] Hand-foot-mouth disease (HFMD) is listed as a Class C infectious disease in my country's "Infectious Disease Prevention and Control Law". It is an acute infectious disease caused by a variety of enteroviruses, and it is more common in infants and young children. . Patients manifested as mouth pain, anorexia, low fever, small herpes or small ulcers on the hands, feet, mouth and other parts. Most patients healed themselves in about a week, and a few patients could cause myocarditis, pulmonary edema, aseptic meningoencephalitis, etc. complication. Individual critically ill children develop rapidly and lead to death. In addition to the more common types of EV71 (Enterovirus 71, EV71) and CA16 (Coxasckievirus A16, CA16), viruses tha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/125A61P31/14
CPCA61K39/12A61K2039/5252A61K2039/545A61K2039/55505A61K2039/70C12N2770/32334A61K2300/00
Inventor 刘可心刘兆梅王巍巍张燕张星星李雅静高强尹卫东
Owner SINOVAC BIOTECH
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