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Construction and application of fusion protein vaccine platform

A fusion protein and vaccine technology, applied in the fields of genetic engineering and biomedicine, can solve problems such as weakened immune system, a large amount of time for vaccines, and problems in the supply of chicken embryos

Pending Publication Date: 2022-01-04
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even though currently approved influenza vaccines confer good protection in healthy young adults, there are several issues that need to be addressed
For example, the production route of some vaccines depends on chicken embryos, such as fire-extinguished influenza vaccines and attenuated influenza vaccines. A disadvantage of these vaccines is that if the prevailing virus strains are of poultry origin, at the same time, the epidemic of the disease will lead to the separation of vaccines and chicken embryos. Increased demand, which can lead to problems with the supply of chicken embryos (6)
Another downside is the enormous amount of time these vaccines take to create
Older adults are more prone to severe syndromes of influenza viruses, and standard vaccines are generally less effective in older adults, whose immune systems weaken with age (7)

Method used

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  • Construction and application of fusion protein vaccine platform
  • Construction and application of fusion protein vaccine platform
  • Construction and application of fusion protein vaccine platform

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0235] Embodiment 1. Design of vaccine platform

[0236] The vaccine platform of interferon-target antigen-immunoglobulin Fc (or antibody) structural unit is composed of three structural units, the first structural unit is the interferon part, and the second structural unit is the immunoglobulin Fc region (or antibody) , the third unit is the target antigen. In actual construction, the three structural units can be arranged and combined in any form and the target antigen can be connected to the Th cell helper epitope through the linker sequence 2. Its representative form is as follows:

[0237] figure 1 . A schematic diagram of the arrangement and combination of the vaccine platform in the form of homologous dimers according to the order of interferon-connecting fragment 1-target antigen-immunoglobulin Fc.

[0238] figure 2 .In the form of heterodimers for the vaccine platform, according to the schematic diagram of the combination of interferon-connecting fragment 1-IgG1-...

Embodiment 2

[0244] Example 2. Construction, purification and production of the vaccine platform

[0245] We describe the expression and production of this vaccine platform by taking the homodimer form of hepatitis B virus Pres1 and coronavirus SARS-CoV-2 RBD protein as examples.

[0246] 1. Vector construction, host cell transfection and induced expression

[0247] 1.1. Using PEE12.4 as the carrier, construct the vaccine structural unit on the carrier by means of molecular cloning, so as to obtain a plasmid that can express the fusion protein, and then transiently transfect 293F cells, collect the culture supernatant, and finally pass ProteinA affinity Chromatographic column purification of the target protein.

[0248] Vector construction (taking HBV preS1 antigen as an example)

[0249] (1) PEE12.4-HindIII-signal peptide 1-interferon-BsiwI-Pres1-BstbI-hIgG1-EcoRI

[0250] (2) PEE12.4-HindIII-signal peptide 1-interferon

[0251] -BsiwI-RBD(SARS-CoV-2)-BstbI-hIgG1-EcoRI

[0252] (3) P...

Embodiment 3

[0282] Example 3. IFNα-Pres1-Fc, Pres1-Fc can induce a stronger immune response in mice than pure Pres1 antigen.

[0283] Materials: C57BL / 6 male mice (5-8) weeks were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.; horseradish peroxidase (HRP)-labeled goat anti-mouse IgG was purchased from Beijing Kangwei Biology Technology Co., Ltd.; 96-well ELISA assay plate was purchased from Corning Costar Company; ELISA chromogenic solution was purchased from eBioscience Company; the microplate reader SPECTRA max PLUS 384 used was purchased from Molecular Company of the United States. The aluminum adjuvant used was purchased from SIGMA Company.

[0284] method:

[0285] (1) Pres1 fusion protein immunized mice, 80 pmol IFN-Pres1-Fc or 80 pmol Pres1-Fc, Pres1 protein was mixed with aluminum adjuvant and subcutaneously immunized mice. At the designated time points, the serum of the mice was collected by taking blood from the orbit for antibody detection.

...

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Abstract

The invention relates to construction and application of a fusion protein vaccine platform. The present invention provides a vaccine, which comprises a fusion protein containing an interferon-target antigen-immunoglobulin Fc region (or antibody) and a Th cell helper epitope. The invention also relates to the use of a fusion protein containing an interferon-target antigen-immunoglobulin Fc (or antibody) region and a Th cell helper epitope for the preparation of a prophylactic or therapeutic composition. The vaccine provided by the invention can be generated through an eukaryotic cell expression system, the vaccine can be inoculated through subcutaneous / muscle or nasal cavity immune ways and the like, and can cause strong immune response of a body. The vaccine of the present invention can be used as a prophylactic or therapeutic vaccine.

Description

technical field [0001] The invention belongs to the technical fields of genetic engineering and biomedicine, and specifically relates to a vaccine, for example, a vaccine comprising a fusion protein containing interferon-target antigen-immunoglobulin Fc region (antibody) as the main framework. The vaccine of the present invention can be used as a vaccine platform for preventing hepatitis B virus (HBV) infection, HPV, EBV, HIV, SARA-COV2, influenza virus infection and HPV, the occurrence of EBV-related tumors and treatment of chronic hepatitis B (chronic hepatitis B , CHB) infection, treatment of HBV, HP, EBV related tumor purposes. Background technique [0002] There are about 257 million chronic viral infections in the world, and about 88,700 people die each year from end-stage liver disease caused by HBV, including liver failure, liver cirrhosis, and hepatocellular carcinoma (1-3). About 30% of patients with liver cirrhosis About 40% of hepatocellular carcinomas (HCC) are...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/12A61K39/39A61K38/21A61K47/68A61P31/04A61P31/12A61P35/00C07K14/005C07K14/555C07K14/56
CPCA61K39/0011A61K39/12A61K39/39A61K47/68A61K38/21C07K14/555C07K14/56C07K14/005A61P35/00A61P31/12A61P31/04A61K2039/575A61K2039/57C07K2319/30C07K2319/02C07K2319/00C12N2730/10122C12N2730/10134C12N2770/20022C12N2770/20034
Inventor 彭华孙士玉其他发明人请求不公开姓名
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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