Respiratory syncytial virosome vaccine and preparation method thereof

A syncytial virus, respiratory technology, used in antiviral agents, pharmaceutical formulations, antibody medical ingredients, etc.

Inactive Publication Date: 2011-06-01
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

There is currently no successful RS Virosome vaccine, so the development of RS Virosome is an extremely potential vaccine development

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Screening and Determination of Respiratory Syncytial Virus Vaccine Strains

[0045] Clinical throat swab samples of respiratory tract infection were collected from 20 provinces and cities CDC in different regions of my country, east, west, north, south, middle, and the RSV virus was amplified and purified by cell culture for vaccine production. Tests such as biology, virus titration and exogenous factors, etc., to screen candidate dominant vaccine virus strains. Further prepare the immune serum of candidate dominant vaccine virus strains, and carry out immunization with clinically isolated RSV epidemic wild strains in different regions in recent years, purchased A-type RSV A2 standard strains, B-type G8537 standard strains, and RSV clinically isolated strains preserved by China CDC and China Inspection Institute. Cross-neutralization studies, the results show that the immunoneutralization crossover between the identified candidate RSV type A and B vaccine stra...

Embodiment 2

[0046] Example 2: Cell culture and cell batch bank establishment

[0047] Take human diploid cells (2BS Cell) as an example: 2BS cells come from American Type Culture Collection (ATCC). The culture medium adopts Dulbecco's modified Eagle medium (DMEM), adds gentamicin, does not add or adds a certain amount of fetal calf serum (2-10%) to cultivate in cell factory or cell fermenter, establishes cell work seed batch library, and And the foreign aid source factors, tumorigenicity and stability after passage are comprehensively tested, and the number of cell generations used is controlled within 40 generations, which all meet the requirements of vaccine production media.

[0048] The cell culture and cell batch bank establishment can be one of the following cells: ① human diploid cells; ② ELL-0 cells; ③ Vero cells; ④ CHO cells; ⑤ Hep-2 cells; ⑥ MDCK cells.

Embodiment 3

[0049] Example 3: Large-scale production and purification of RSV culture and proliferation

[0050] ① RSV was inoculated in human diploid cells (2BS) cells covering the bottom of the flask, cultured at 37°C for 48 hours, collected cells, and centrifuged at 5000rmp at 4°C for 5 minutes to remove cells and impurities. ②The supernatant was taken and subjected to sucrose gradient centrifugation. The sucrose was prepared with PBS buffer, and the system used 4 mL of 30% sucrose; 4 mL of 45% sucrose; 1 mL of 60% sucrose. Centrifuge at 100000g for 2h at 4°C. ③ Aspirate 30% and 40% interlayers, add an appropriate amount of PBS to dilute, and continue ultracentrifugation at 100,000 g at 4°C for 2 hours. ④ Collect the precipitate and measure the density with a refractometer. Store in RSV preservation solution (17.4% Glycerol, 145mM NaCl, 2.5mM HEPES, 0.1mM MgCl2, 0.1mM CaCl2, pH 7.4) for later use. The RSV includes A subtype and B subtype.

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Abstract

The invention discloses a respiratory syncytial virosome vaccine, which mainly comprises a respiratory syncytial virosome, wherein the virosome comprises a complete respiratory syncytial virus (RSV) surface, and can stimulate good dual cell and body fluid immune response. The virosome protein antigens are prepared into formulations such as injection, nasal spray, sublingual tablet and transdermal agent by adding or not adding an adjuvant, can safely and effectively prevent RSV infection after being immunologically inoculated to different animals or people, and provide the ideal vaccine for the safe and effective immune prevention and control of the RSV infection of different age groups. Researches show that the respiratory syncytial virosome vaccine has a good immunological protection effect on the different age groups.

Description

technical field [0001] The invention relates to a respiratory syncytial virus vaccine and a preparation method thereof, in particular to a virosome vaccine with a complete surface antigen of the respiratory syncytial virus and a preparation method thereof. Background technique [0002] Respiratory syncytial virus (RSV) is one of the main causes of lower respiratory tract infection in infants and young children worldwide. RSV infection can cause a series of respiratory diseases, ranging from simple cold symptoms to severe lower respiratory tract infections. Infection and complications. The first infection of RSV mostly occurs in 2-month-old infants, the infection rate is as high as 83%, and no lasting immunity will be produced after infection, and 50% of infants will be re-infected 1 to 2 years after the first infection. At the same time, RSV is also a common and important pathogen in adolescents, the elderly, and immunocompromised populations. RSV belongs to the family Par...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/155A61K39/39A61K39/295A61P31/14C12N7/02
Inventor 王希良曹政杨鹏辉王鋮高啸赵忠鹏罗德炎段跃强邢丽
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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