Nanoparticle containing EV71VP1 protein and preparation method of nanoparticle

A nanoparticle and protein technology, applied in the field of medicine, can solve problems such as increased eosinophils, inability to activate cellular immunity, and sterile abscesses

Active Publication Date: 2017-02-22
GUANGZHOU LIDE BIOMEDICINE TECH CO LTD
View PDF2 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Aluminum adjuvant is the only adjuvant approved for human use, but many studies have shown that aluminum adjuvant can only activate humoral immunity but not cellular immunity
Another study reported that aluminum adjuvant can cause aseptic abscess, eosinophilia, fasciitis and other side effects

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Nanoparticle containing EV71VP1 protein and preparation method of nanoparticle
  • Nanoparticle containing EV71VP1 protein and preparation method of nanoparticle
  • Nanoparticle containing EV71VP1 protein and preparation method of nanoparticle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0112] The preparation of embodiment 1 nanoparticle

[0113] 1. Reagents: EV71 VP1 protein is commercially available (produced by Escherichia coli expression system, then dissolved and refolded), and its amino acid sequence is shown in SEQ ID NO:1.

[0114] CpG ODN1826 is commercially available, and its nucleotide sequence is shown in SEQ ID NO:2.

[0115] All other reagents were purchased commercially.

[0116] 2. Preparation process:

[0117] (1) Disperse chitosan (molecular weight 47KDa, degree of deacetylation 90%) in sterilized distilled water, under magnetic stirring, add acetic acid with a volume fraction of 1%, stir overnight, and filter with filter paper to obtain shell Polycan solution, chitosan concentration is 0.45mg / mL.

[0118] (2) Disperse the heparin in sterilized distilled water, stir for 5 minutes under magnetic stirring, and filter through a 0.22 μm filter membrane to obtain a heparin solution with a heparin concentration of 0.3 mg / mL.

[0119] (3) The E...

Embodiment 2

[0127] Embodiment 2. Morphological characterization, particle size test and potential test of the nanoparticles:

[0128] 1. Morphological characterization:

[0129] The chitosan-heparin nanoparticles wrapped with EV71 VP1 protein and CpG, and the chitosan-heparin nanoparticles wrapped with EV71 VP1 protein and TNF-α were observed by transmission electron microscopy. The morphology of the two nanoparticles is as follows image 3 A and 3B are shown.

[0130] As shown in the figure, the two kinds of nanoparticles have regular shape, round shape, smooth surface, good dispersion, and no obvious adhesion, damage, collapse and other phenomena.

[0131] 2. Particle size test and Zeta potential test:

[0132] Chitosan-heparin nanoparticles coated with EV71 VP1 protein and CpG, and chitosan-heparin nanoparticles coated with EV71 VP1 protein and TNF-α were averaged using a Malvern particle sizer (with a dynamic light scattering detector). The particle size and Zeta potential were tes...

Embodiment 3

[0136] Calculation of Encapsulation Efficiency of EV71 VP1 Protein and Immunological Adjuvant in Example 3 Nanoparticles

[0137] 1. Take the CS+VP1+CpG+Heparin nanoparticle solution, centrifuge at 4°C and 60,000g for 20 minutes, take the supernatant, record the volume, and use the EV71 VP1 Elisa kit to detect the content of free EV71 VP1 protein in the supernatant. The encapsulation efficiency of EV71 VP1 protein in nanoparticles was calculated according to the following formula.

[0138] Encapsulation efficiency of EV71 VP1 protein=w 0 -w 1 / w 0 ×100%, where w 0 is the total amount of EV71 VP1 protein added; w 1 is the total amount of free EV71 VP1 protein in the supernatant.

[0139] 2. Take the CS+VP1+CpG+Heparin nanoparticle solution, centrifuge at 4°C and 60,000g for 20 minutes, take the supernatant, record the volume, use a Nanodrop spectrophotometer to detect the content of free CpG in the supernatant, and calculate according to the following formula Encapsulatio...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
electric potential / voltageaaaaaaaaaa
sizeaaaaaaaaaa
Login to view more

Abstract

The invention belongs to the technical field of medicine, and relates to a nanoparticle containing EV71VP1 protein and a preparation method of the nanoparticle. Specifically, the invention relates to a nanoparticle containing EV71 VP1 protein, an immunologic adjuvant, a cationic polymer and an anionic polymer, wherein the immunologic adjuvant is selected from one or more of TNF-alpha, CpG, bcg vaccine and flagellin. The invention further relates to a method for preparing the nanoparticle including immunogenic composition (such as vaccine) of the nanoparticle. The nanoparticle is used for preparing the immunogenic composition (such as vaccine) and for the nanoparticle or the immunogenic composition (such as vaccine) to initiate or enhance the immune response of subjects to EV71.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a nanoparticle comprising VP1 protein of EV71 virus, an immune adjuvant, a cationic polymer and an anionic polymer, and the immune adjuvant is selected from TNF-α, CpG, BCG and flagellin one or more of. The invention also relates to methods for the preparation of said nanoparticles, immunogenic compositions (such as vaccines) comprising said nanoparticles, the use of said nanoparticles for the preparation of immunogenic compositions (such as vaccines), and said Use of nanoparticles or an immunogenic composition (such as a vaccine) to elicit or enhance an immune response to EV71 in a subject. Background technique [0002] HFMD is a global infectious disease caused by a variety of enteroviruses, the main pathogens of which are Coxsackie virus (CV, CV) and Echo virus (ECHO virus, EO) and Enterovirus 71 (EV71). Among them, CV16 and EV71 infections are the most common. HFMD mainly ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/125A61K39/39A61K9/51A61K47/36A61P31/14
CPCA61K39/12A61K39/39A61K9/5161A61K2039/55561A61K2039/55522C12N2770/32334
Inventor 刘利新乔冬冬陈永明
Owner GUANGZHOU LIDE BIOMEDICINE TECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap