Respiratory syncytial virus-like particle vaccine and preparation method thereof

A technology for syncytial virus and vaccine, which can be used in antiviral agents, pharmaceutical formulations, medical preparations containing active ingredients, etc., and can solve the problems of difficulty in large-scale preparation and low VLP formation efficiency.

Inactive Publication Date: 2011-04-27
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

Existing studies have shown that the VLP formation mechanism of RSV is unknown so far. Some scholars have used RSV protein to study VLP formation, and the results show that the VLP formation efficiency of its own protein is low, and it is difficult to achieve large-scale production.

Method used

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  • Respiratory syncytial virus-like particle vaccine and preparation method thereof
  • Respiratory syncytial virus-like particle vaccine and preparation method thereof
  • Respiratory syncytial virus-like particle vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1: The gene and protein composition of respiratory syncytial virus-like particle vaccine of the present invention

[0063] Take the nucleic acid and protein sequence of M protein of RSV, the nucleic acid and protein sequence of NP protein of RSV, the nucleic acid and protein sequence of F protein of RSV, the nucleic acid and protein sequence of G protein of RSV, the nucleic acid sequence and protein sequence of M protein of NDV, The nucleic acid sequence and protein sequence of the NP protein of NDV, the nucleic acid sequence and protein sequence of the HN protein of NDV, and the nucleic acid sequence and protein sequence of the F protein of NDV are shown in Table 1-20.

Embodiment 2

[0064] Embodiment 2. The expression vector construction of respiratory syncytial virus-like particle vaccine of the present invention

[0065] Take the construction of recombinant pHWD2000 expression vector as an example: use PCR to amplify the RSV M gene white nucleic acid sequence, F protein gene sequence, G protein gene sequence, and NP protein gene sequence and insert them into the pHWD2000 vector to form recombinant pHWD2000-M, pHWD2000 -F, pHWD2000-G, pHWD2000-NP expression vectors. In another scheme, the NDVM gene white nucleic acid sequence, F protein gene sequence, and NP protein gene sequence are inserted into the pHWD2000 vector to form recombinant pHWD2000-M, pHWD2000-F, and pHWD2000-NP expression vectors.

[0066] pHWD2000-NH / G expression vector construction: through bioinformatics methods, analyze the intracellular region (CT) and transmembrane region (TM) of NDV HN protein gene sequence, and analyze the extracellular region of RSV G protein gene sequence at the ...

Embodiment 3

[0068] Example 3. Cell culture and cell batch bank establishment

[0069] Take UMNSAH / DF-1 cell culture and bank building as an example: UMNSAH / DF-1 cells come from American Type Culture Collection (ATCC). The culture medium adopts Dulbecco's modified Eagle medium (DMEM), adds penicillin and streptomycin, does not add or adds a certain amount of fetal bovine serum (10-2%) to cultivate in cell factories or cell fermenters, and establishes cell working seed batch banks. In addition, the foreign aid source factors, tumorigenicity and stability after passage were comprehensively tested, and the number of cell generations used was controlled within 60 generations, which all met the requirements of vaccine production media.

[0070] The cell culture and batch cell bank establishment include but not limited to: ① UMNSAH / DF-1 cells; ② human diploid cells; ③ Vero cells; ④ CHO cells; ⑤ Hep-2 cells; ⑥ MDCK cells.

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Abstract

The invention discloses a virus-like particle vaccine (NDV / RSV VLP vaccine) of which the centre is from NVD (Newcastle disease virus) and the surface protein is from RSV (respiratory syncytial virus), and also discloses a preparation method of the virus-like particle vaccine. The RSV VLP vaccine provided by the invention comprises a VLP composed of four structural proteins of respiratory syncytial virus M, F, NP and G; and the NDV / RSV VLP vaccine comprises four structural proteins of Newcastle disease virus M, F, NP and NH and VLP, wherein the VLP is formed by two surface proteins which are a NDV F / RSV F fusion protein composed of Newcastle disease virus F protein and respiratory syncytial virus F protein and an NDV HN / RSV G fusion protein composed of Newcastle disease virus HN protein and respiratory syncytial virus G protein. The test of pesticide effectiveness shows that RSV infection can be safely and effectively prevented after various dosage forms prepared by the VLP protein antigen formed with the method by adding or not adding adjuvants are used for processing immunization for different animals or the crowd, and the vaccine for processing immune prevention for the RSV inflection is supplied for the crowd with different ages.

Description

field of invention [0001] The invention belongs to the field of biopharmaceuticals, relates to a respiratory syncytial virus-like particle vaccine (RS VLP vaccine) and a preparation method thereof, in particular to a virus-like particle vaccine (NDV) whose core comes from Newcastle disease virus and whose surface protein comes from syncytial virus / RSV VLP vaccine) and its preparation method. 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/155C12N15/79A61P31/14
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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