Vaccine based on ferritin nanoparticle, and preparation method thereof

A nanoparticle and ferritin technology, applied in biochemical equipment and methods, medical preparations with non-active ingredients, and medical preparations containing active ingredients, etc. drug properties and other issues to prevent HBV infection, enhance antibody levels, and achieve the effects of efficient production

Active Publication Date: 2018-08-24
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows for quick creation of small particles called nanoparticles containing iron oxide with high affinity towards certain proteins found on infected cells like HepG2 cell membranes. These tiny particles are then attached to magnetic microparticle carriers which help transport them through blood vessels into targeted areas where they stimulate immune responses against these pathogens. Additionally, this technology could have potential applications in treating diseases such as liver cirrhosis caused due to excessive alcohol consumption.

Problems solved by technology

This patented technical problem addressed in this patents relates to improving the stability and activity of proteins involved in viruses like honeycomb shaped particles called FeaNPs, while maintaining their original structure and function during manufacturing processes. Previous attempts were made towards modifying these structures but did not fully address all aspects mentioned above.

Method used

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  • Vaccine based on ferritin nanoparticle, and preparation method thereof
  • Vaccine based on ferritin nanoparticle, and preparation method thereof
  • Vaccine based on ferritin nanoparticle, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1. Construction and production of ferritin nano-vaccine platform

[0032] Materials: 1. Strains: DH5αE.coli, BL21(DE3)PLySs E.coli competent cells were purchased from Beijing Quanshijin Biotechnology Co., Ltd. 2. Plasmid: The pDEST14-SpyCatcher plasmid (containing the full-length gene sequence of SpyCatcher) was donated by Dr. Mark Howarth (University of Oxford, UK) and preserved in our laboratory (Zakeri, Fierer et al. 2012). The pDEST14-ΔN SpyCatcher plasmid was preserved by our laboratory (Liu, Zhou et al. 2014). 3. Reagents: DNA Agarose Gel Recovery Kit, Plasmid Small and Large Plasmid Extraction Kit, DNAMarker, Taq PCR Mastermix and Pfu PCR Mastermix used in molecular biology experiments were purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.; restriction endonuclease and T4 ligase were purchased from New England Biolabs; LB medium, antibiotics (Ampicillin) and protein expression inducer IPTG used for bacterial culture were purchased from...

Embodiment 2

[0052] Example 2. Purification and Characterization of Ferritin Nano Vaccine Platform

[0053] Materials: Superose6Increase 10 / 300 gel filtration prepacked column was purchased from GE HealthCare. 50kDa ultrafiltration tubes were purchased from Merck Millipore. 0.22 μm, 0.45 μm filters were purchased from Pall Corporation. Transmission electron microscope Tecnai Spirit (120kV) manufacturer US FEI. Commonly used chemical reagents (SDS-PAGE gel configuration and electrophoresis buffer, glycine, Tris, Coomassie brilliant blue, etc.) were purchased from Beijing Dingguo Changsheng Biotechnology Co., Ltd. and Amresco.

[0054] method:

[0055] 1. Crude purification Put the ΔN SpyCatcher / SpyTag-ferritin crude extract in a 70°C water bath and heat it for 15 minutes to denature and precipitate most of the miscellaneous proteins that are not resistant to high temperatures. Filter the membrane to remove tiny precipitates to obtain ΔN SpyCatcher / SpyTag-ferritin crude liquid. Concentr...

Embodiment 3

[0059] Embodiment 3, ferritin-preS1, ferritin-M2e, ferritin-HCV pep designed based on ferritin nano-vaccine platform (412-423) , Production, purification and biochemical characterization of ferritin-E7 nanovaccine

[0060] Materials: The plasmid containing the nucleic acid sequence of HBV envelope protein preS1 was donated by the Fu Yangxin Group of the Institute of Biophysics, Chinese Academy of Sciences. It contains the nucleic acid sequence of influenza virus membrane protein M2e, human papillomavirus type 16 E7 protein nucleic acid sequence and fluorescent indicator protein eGFP nucleic acid sequence The plasmid was preserved by this experiment; the 412-423 amino acids of the conserved segment of the hepatitis C virus E2 protein with SpyTag (SEQ ID NO: 17) were synthesized by Jill Biochemical (Shanghai) Co., Ltd.; the Ni-Agarose used for protein purification The His-tag protein purification kit was purchased from Beijing Kangwei Century Biotechnology Co., Ltd.; other strai...

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Abstract

The invention provides a ferritin nanoparticle carrier for preparing a vaccine, and also provides a vaccine based on the ferritin nanoparticle carrier, and a preparation method thereof. The ferritin nanoparticle carrier is formed by self-assembling ferritin with fuse-expressed SpyCatcher, or is formed by self-assembling ferritin with fuse-expressed SpyTag, the vaccine further comprises an antigenwith fuse-expressed SpyTag when the nanoparticle carrier is formed by self-assembling the ferritin with fuse-expressed SpyCatcher, and the vaccine further comprises an antigen with fuse-expressed SpyCatcher when the nanoparticle carrier is formed by self-assembling the ferritin with fuse-expressed SpyTag, wherein the ferritin is covalently linked to the antigen by a SpyCatcher-SpyTag interaction.

Description

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Claims

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Application Information

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Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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