ZIKV (zika virus) vaccine and preparation method thereof

A Zika virus and vaccine technology, applied in the field of vaccines, to achieve the effect of improving infectivity, good virus infection and transmission, and strong immunogenicity

Inactive Publication Date: 2018-06-29
GUANGZHOU N BIOMED LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No Zika virus vaccine candidate currently has NS1 as the primary antigenic target, although reports suggest that NS1-specific immune responses may be protective against Flaviviridae infection

Method used

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  • ZIKV (zika virus) vaccine and preparation method thereof
  • ZIKV (zika virus) vaccine and preparation method thereof
  • ZIKV (zika virus) vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Zika virus vaccine and its preparation method

[0044] In an embodiment of the Zika virus vaccine of the present invention, the Zika virus vaccine is a gene vector capable of expressing Zika virus membrane protein prM / M, envelope protein E and non-structural protein NS1, wherein the vector is a shuttle Plasmid pGA1; the vector contains the coding sequence of Japanese encephalitis virus signal peptide, the coding sequence of membrane protein prM / M, the coding sequence of envelope protein E, the 2A self-splicing sequence and the recombination of the coding sequence of non-structural protein NS1 in sequence The recombinant sequence is inserted into the open reading frame of the non-structural protein NS1, and the coding sequences of the membrane protein prM / M, the envelope protein E and the non-structural protein NS1 are optimized according to the mammalian codon preference.

[0045] The method for preparing the Zika virus vaccine includes the following steps:

[0046]...

Embodiment 2

[0048] Example 2 Construction of a recombinant adenovirus vector carrying the coding sequence of the Zika virus main antigen

[0049] 1. Recombinant adenovirus vector plasmid construction.

[0050] Zika virus E protein, prM / M protein, and NS1 protein sequences are from Zika virus isolate 1_0080_PF (GenBank No: ANO46313.1). The coding gene sequences of these proteins were optimized based on mammalian cell codons and obtained through total gene synthesis (GenScript, China). The fusion gene sequence Ad2-prM / E-NS1 consists of (5' end to 3'end): JEV (Japanese Encephalitis Virus) signal peptide, prM / M, E, 2A self-splicing sequence, NS1; Ad2-prM / The map of E-NS1 sequence is as figure 1 As shown, the base sequence of the fused prM / E-NS1 is shown in SEQ ID NO. 1, and the corresponding amino acid sequence is shown in SEQ ID NO.2.

[0051] In order to compare the beneficial effects of the recombinant Zika virus containing the NS1 antigen, this example also constructed a recombinant adenoviru...

Embodiment 3

[0054] Example 3 Identification of antigen expression of recombinant adenovirus vector Zika virus vaccine

[0055] Vero cells were infected with Ad2-E, Ad2-prM / E, Ad2-prM / E-NS1 and Ad2-empty empty vector controls respectively, and the infection dose was 100 viral particles (vp) per cell. 48 hours after infection , Collect infected cells and culture supernatant. Western-blot was used to detect the expression of antigen protein. The protein samples were electrophoresed by SDS-PAGE gel, transferred to PVDF membrane, blocked with PBST containing 5% skimmed milk powder for 1 hour at room temperature, and then incubated with anti-Zika virus E protein antibody or anti-Zika virus NS1 protein antibody at room temperature for 1 hour . Then, the goat anti-human secondary antibody labeled with horseradish peroxidase was incubated. Finally, the horseradish peroxidase substrate was used for color development.

[0056] Experimental results: such as figure 2 As shown, Ad2-E, Ad2-prM / E, Ad2-pr...

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PUM

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Abstract

The invention discloses a ZIKV (zika virus) vaccine. The ZIKV vaccine contains or can express membrane protein prM/M, envelope protein E and non-structural protein NS1 of ZIKV. The invention also discloses a preparation method of the ZIKV vaccine. The ZIKV vaccine has higher immunogenicity and can more effectively prevent ZIKV infection.

Description

Technical field [0001] The invention relates to the technical field of vaccines, in particular to a Zika virus vaccine and a preparation method thereof. Background technique [0002] Zika Virus (ZIKV) belongs to the Flaviviridae family and is a single-stranded positive-stranded RNA virus. Historically, Zika virus was only circulating in a small area in tropical regions of Africa and Asia, and symptoms were mild after infecting healthy adults. Since 2007, Zika virus has continued to circulate in Latin America. In these epidemics, Zika virus infection can cause serious neurological diseases, including microcephaly and Guillain-Barre syndrome. What is more noteworthy is that Zika virus can be transmitted not only through mosquito vectors, but also through sexual contact. The Zika virus epidemic has brought serious health threats and economic losses to people in the affected areas. At present, there is no clinically effective anti-Zika virus drug, and there is no approved Zika vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/14C12N15/861C12N15/40
CPCA61K39/12A61K2039/5256A61K2039/53A61K2039/70C12N15/86C12N2710/10343C12N2770/24134C12N2800/107C12N2800/22A61K2300/00Y02A50/30
Inventor 陈凌冯立强
Owner GUANGZHOU N BIOMED LTD
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