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Intradermal injection technology for enhancing viral antigen immunogenicity and application of intradermal injection technology in research of hand-foot-mouth disease vaccine

A technology for hand, foot and mouth disease and vaccines, applied in the direction of virus antigen components, vaccines, viruses, etc., can solve the problems of complex etiology, limited cross-protection ability, etc., and achieve good growth and replication, good safety, and improved quality.

Active Publication Date: 2019-04-12
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although so far, research on antiviral drugs and corresponding vaccines has achieved some results, but with the use of inactivated vaccines against EVA71, changes in the pathogenic spectrum of hand, foot and mouth disease, and the cross-protective ability of various enteroviruses The research of multivalent HFMD preventive vaccines against CVA16 and other enteroviruses has become a very urgent problem
However, under such a background of complex etiology and unclear pathogenic mechanism of infection, vaccine development and exploration are carried out, especially in the case of many technical bottlenecks encountered in the development of CVA16 vaccine, multivalent vaccine for hand, foot and mouth disease The establishment of the technical process puts forward higher requirements

Method used

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  • Intradermal injection technology for enhancing viral antigen immunogenicity and application of intradermal injection technology in research of hand-foot-mouth disease vaccine
  • Intradermal injection technology for enhancing viral antigen immunogenicity and application of intradermal injection technology in research of hand-foot-mouth disease vaccine
  • Intradermal injection technology for enhancing viral antigen immunogenicity and application of intradermal injection technology in research of hand-foot-mouth disease vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The present embodiment provides a kind of preparation method of bivalent hand, foot and mouth disease inactivated vaccine for human use, and the specific steps are as follows:

[0042] (1) Cell culture: According to the KMB-17 cell production and verification regulations, select the 28th passage KMB-17 cells with dense growth, discard the culture medium, digest with 0.1% trypsin, discard the trypsin solution, and use the original culture medium volume 4% volume of DMEM to suspend the cells to obtain a cell suspension;

[0043] (2) Virus inoculation and culture: Add an appropriate amount of the above cell suspension into the cell factory, mix well and place at 37°C for 3-4 days; after growing into a dense monolayer, discard the culture medium; add EVA71 or CVA16 virus seeds respectively , the moi of the inoculum is 0.02-0.05; after mixing, place it at 37°C and shake it gently at the right time; after 30 minutes, add quantitative cell growth medium and place it at 37°C fo...

Embodiment 2

[0049] This example is the distribution detection of various intermediate products during the preparation process of the human bivalent inactivated hand, foot and mouth disease vaccine described in Example 1.

[0050] Among them, the virus harvest liquid, virus concentrated liquid, inactivated and purified antigen liquid, and semi-finished products are all clear liquids without foreign matter and precipitation. The sterility test and mycoplasma test are all negative, and there is no abnormal toxin, pH 7.2-8.0, see Table 1 .

[0051]

[0052] Such as figure 1 As shown, samples were taken at different time points after the virus was inoculated into KMB17 cells, and the infectious titer was detected. The results show that initial typical lesions can appear within 8 hours of inoculating cells, and the peak of lesions can appear within 24-48 hours, and the titer is usually 7.0±0.5LogCCID 50 .

Embodiment 3

[0054] This example tests the safety of the finished product of the bivalent inactivated vaccine for human use prepared in Example 1.

[0055] The finished product of the vaccine was injected at 0.2 mL / only into the abdominal cavity, intradermal, and muscle of mice with a body weight of 18-20 g. There were 20 mice in each group. There should be no death within one month. The weight gain of the mice was the same as that of the normal saline control group, see Table 2.

[0056]

[0057] The formaldehyde content of the finished vaccine is not higher than 50μg / dose, and the bacterial endotoxin is not higher than 100EU / dose. And it was negative in the sterility test, see Table 3.

[0058]

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Abstract

The invention provides an intradermal injection technology for enhancing viral antigen immunogenicity and an application of the intradermal injection technology in research of a hand-foot-mouth disease vaccine. The intradermal injection technology is applicable to the human divalent hand-foot-mouth disease vaccine prepared from EVA71 (enterovirus 71) and CVA16 (coxsachievirus 16). The vaccine takes diploid fibroblasts (KMB17 cells) derived from human embryo lung as a passage cell matrix. Through comparison of intradermal and intramuscular injection of the human divalent hand-foot-mouth diseasevaccine, results show that the intradermal injection technology can more rapidly stimulate innate immunity system related components, enhance the viral antigen immunogen effect, induce and activate natural immunoreactions, start specific immunoreactions and effectively resist viral pathopoiesis.

Description

technical field [0001] The invention belongs to the technical field of human preventive medicine. More specifically, the invention relates to an immunization injection technique for enhancing virus antigen immunity, which is suitable for inactivated vaccines for human hand, foot and mouth disease. Background technique [0002] Human hand, foot and mouth disease (HFMD) caused by enterovirus type 71 (EVA71) and coxsackievirus A group 16 (CVA16) infection has attracted special attention from public health management in the Asia-Pacific region in recent years. One of the viral infectious diseases, the main reason why this infectious disease has received widespread attention includes two aspects: First, it has caused several large-scale outbreaks involving a large number of people and affecting a wider area in various regions of the world. Second, in the infected population, a small number of infected individuals can be found to have severe nervous system damage and clinical fea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/125A61P31/14C12N5/077C12N7/00C12R1/93
CPCA61K39/12A61P31/14C12N5/0656C12N7/00A61K2039/54A61K2039/5252A61K2039/70C12N2770/32334C12N2770/32351Y02A50/30
Inventor 李琦涵范胜涛张莹徐兴丽冯敏董承红杨二霞蒋国润廖芸姜莉王丽春廉亚茹崔萍芳
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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