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Respiratory syncytial virus pre-fusion F protein and application thereof

A technology of syncytial virus and respiratory tract, applied in the pre-fusion F protein of respiratory syncytial virus and its application field, achieving good protection and structural stability

Active Publication Date: 2019-07-26
BEIJING JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although, since 1956, RSV has been widely recognized, but so far there is no effective vaccine against RSV infection.

Method used

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  • Respiratory syncytial virus pre-fusion F protein and application thereof
  • Respiratory syncytial virus pre-fusion F protein and application thereof
  • Respiratory syncytial virus pre-fusion F protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0039] Experiment 1: Construction of a recombinant plasmid capable of expressing F protein with a mutation in the furin cleavage site.

[0040]Based on the optimized F protein gene of the RSV Long strain, we mutated its furin cleavage site. The amino acid sequence and DNA sequence of each mutant are shown in Table 1, where the bold font is the furin cleavage site. F is the genome-optimized RSV F protein, M1-F is the mutant with the first furin cleavage site mutated, M2-F is the mutant with the second furin cleavage site mutated, U-F is the mutant without Cleavage mutants, that is, mutants in which two sites are mutated at the same time.

[0041] Table 1:

[0042]

[0043]

[0044] In addition, this experiment constructed a mutant DS-Cav1 in the pre-fusion structure as a control, but the C-terminus of F1 retains the transmembrane region (transmembrane region, TM region) and intracellular region (cytoplasmic region CT region) of F1 structure. In the experiment, the M1-F...

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Abstract

The invention provides a respiratory syncytial virus pre-fusion F protein and application thereof, and belongs to the technical field of respiratory syncytial virus vaccines. The pre-fusion F proteinis formed in the way that the 106th, 108th and 109th amino acids of a wild-type F protein are mutated from Arg to Asn, the 104th amino acid is mutated from Asn to Cys, the 155th amino acid is mutatedfrom Ser to Cys, or the 58th amino acid is mutated from Thr to Cys, and the 190th amino acid is mutated from Ser to Cys. A first furin cleavage site of the mutant wild-type F protein makes the structure of Pre-F more stable, the expression amount is not decreased, and the pre-fusion epitope of M1-F expression in 293T cells is significantly increased. The second mutation of the amino acid site of the F protein forms a disulfide bond, achieves the more stable pre-fusion epitope compared to the wild type, and is suitable for other strains of human RSV. The protein is suitable for various vaccineforms with the RSV F protein as the antigen, such as nucleic acid vaccines, protein vaccines, vector vaccines and recombinant virus particle vaccines.

Description

technical field [0001] The invention relates to the technical field of respiratory syncytial virus vaccines, in particular to a pre-fusion F protein of respiratory syncytial virus and its application. Background technique [0002] Human respiratory syncytial virus (RSV) is an enveloped, nonsegmented, single-stranded, negative-sense RNA virus belonging to the Pneumoviridae family that can cause Severe lower respiratory infection. RSV causes 30 million severe lower respiratory tract infections in children under the age of 5 worldwide each year, and the death toll reaches more than 60,000. RSV infection accounted for 6.7% of all-cause deaths among infants aged 1 month to 1 year. 100% of infants have been infected with RSV virus at least once within 3 years of age, and the peak of infection occurs at 2 to 4 months of age. Children younger than 5 years of age are at risk of serious illness from RSV infection, and recurrent infections with the virus are common. Although, since...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/135C12N15/45G01N33/68G01N33/569C07K16/10A61K39/155A61P31/14
CPCA61K39/12A61K2039/5258A61K2039/53A61P31/14C07K14/005C07K16/1027C12N2760/18522C12N2760/18534G01N33/56983G01N33/68G01N2333/135
Inventor 何金生马尧付远辉彭向雷郑妍鹏
Owner BEIJING JIAOTONG UNIV
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