Univalent and bivalent gene engineered subunit vaccine for hand-foot-and-mouth disease and preparation method thereof

A subunit vaccine, hand, foot and mouth disease technology, applied in the field of biopharmaceuticals, can solve the problems of undeveloped vaccines with commercial application value

Inactive Publication Date: 2010-04-21
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011]Despite the development of EV71 and Cox.A16 vac

Method used

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  • Univalent and bivalent gene engineered subunit vaccine for hand-foot-and-mouth disease and preparation method thereof
  • Univalent and bivalent gene engineered subunit vaccine for hand-foot-and-mouth disease and preparation method thereof
  • Univalent and bivalent gene engineered subunit vaccine for hand-foot-and-mouth disease and preparation method thereof

Examples

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Embodiment 1

[0044] Embodiment 1: Preparation of hand, foot and mouth disease subunit vaccine of the present invention

[0045] 1. Preparation of recombinant baculovirus Bac-EV71-P1-3CD and Bac-Cox.A16-P1-3CD

[0046] Cells: African green monkey kidney cell line (Vero cell line) and insect cell line Sf-9 were used.

[0047] Viruses: H13-Henan-08 was used for strain EV71, and S10-Shandong-08 was used for strain Cox.A16.

[0048] Cell culture: resuscitate the cells, resuscitate the cells in one ampoule into one cell culture flask, after 4-6 days of culture, spread out 4 cell culture flasks according to the split ratio of 1:4.

[0049] Virus inoculation: after cell culture for 3 to 5 days, discard the cell culture supernatant, and inoculate with HFMD virus. The virus inoculation amount is 0.1 MOI, the culture temperature is 37° C., the culture time is 30-40 hours, and the virus maintenance solution is DMEM medium containing 2% fetal bovine serum.

[0050] Harvest virus: When the cells have...

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Abstract

The invention discloses a univalent and bivalent gene engineered subunit vaccine for preventing Enterovirus 71 (EV71) and Coxsackie virus A16 (Cox.A16) of hand-foot-and-mouth disease, and a preparation method thereof. The preparation method comprises the following steps: respectively obtaining recombinant baculovirus Bac-EV71-P1-3CD and Bac-Cox.A16-P1-3CD by gene engineering means, respectively efficiently coexpressing similar SeQ ID No.1 EV71 P1 and Se Q ID No.2 Cox.A16 P1 and 3CD proteins in insect cells, and respectively self-assembling into EV71 VLP and Cox.A16 VLP; establishing and verifying a vaccine strain third-stage seed lot library; culturing cells; inoculating and propagating virus; lysing the cells, ultra-filtering and purifying virus suspension; and further preparing the univalent and bivalent vaccine. The vaccine has good application prospect for preventing the hand-foot-and-mouth disease.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and relates to a vaccine and a preparation method thereof, in particular to a hand-foot-mouth disease single- and bivalent genetic engineering subunit vaccine and a preparation method thereof. technical background [0002] Hand, foot and mouth disease is caused by viruses of the family Picornaviridae and the genus Enterovirus, and has become an important infectious disease that endangers the health of infants and young children and public health safety. HFMD is mainly caused by Enterovirus 71 (EV71) and Coxsackievirus A16 (Cox.A16). The clinical symptoms caused by infection with EV71 and Cox.A16 were indistinguishable. Infection with EV71 is often complicated by severe neuroinflammation, including viral meningitis, encephalitis, and paralysis similar to polio, resulting in pulmonary edema and pulmonary hemorrhage, which often lead to rapid death in infants and young children. Cox.A16 is an imp...

Claims

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Application Information

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IPC IPC(8): A61K39/125A61K39/295A61P31/14C12N15/866C12N7/04C12R1/93
Inventor 赵忠鹏李敏高啸罗德炎李晓楠刘鑫邢丽段越强杨鹏辉王希良
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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