Supercharge Your Innovation With Domain-Expert AI Agents!

Novel virus-like particle of enterovirus 71, its preparation method and application

An enterovirus, virus-like technology used in the field of biomedicine to solve problems such as death

Active Publication Date: 2022-03-29
中国科学院上海免疫与感染研究所
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although EV71 infection usually manifests as a mild, self-limited disease, it can also lead to severe neurological complications, such as brainstem encephalitis, poliomyelitis-like paralysis, and pulmonary edema, and even death

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
  • Novel virus-like particle of enterovirus 71, its preparation method and application
  • Novel virus-like particle of enterovirus 71, its preparation method and application
  • Novel virus-like particle of enterovirus 71, its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0048] In order to reduce the cost of VLP preparation and the risk of clinical application, the inventors truncated different proteins that form VLPs to explore their effects on the assembly and immunogenicity of VLPs. The results of the present inventors show that after truncating amino acids (amino acid, aa) at positions 4-30 of the VP1 protein, VLPs can still assemble, and the size and shape of the particles are similar to those of full-length VLPs. The present inventors evaluated the immunogenicity and in vivo protective effect of VLPs of truncated versions of VP1. The results showed that truncated VLPs induced total IgG responses and neutralizing antibody responses at levels comparable to those of full-length VLPs. Moreover, the maternal immunization / challenge experiment also confirmed that the truncated version of VLPs according to the present invention can provide protection in vivo similar to that of full-length VLPs. Taken together, these results confirmed that VP1 p...

Embodiment 1

[0122] Example 1 Expression and identification of VP1 truncated EV71 VLPs in Pichia pastoris

[0123] The truncated VP1 protein constructed by the inventors includes 4-30aa or 4-73aa truncated VP1 protein respectively. The schematic diagram of the plasmid is as figure 1 As shown in A. Plasmid YE010 was used to co-express 3CD and P1 protein with truncated 4-30aa of VP1 protein, and plasmid YE011 was used to co-express 3CD and P1 protein with truncated 4-73aa of VP1 protein.

[0124] The inventors transformed yeast cells with YE010 and YE011 plasmids respectively, and used yeast cells transformed with empty vectors as negative controls, YE003 (not truncated) [3] Transformed yeast cells were used as positive controls. The obtained recombinant yeast clones were used to measure the EV71 antigen content by ELISA method, and were arranged according to the order of antigen content from high to low. Such as figure 1 As shown in B, no EV71 antigen was detected in the yeast transfor...

Embodiment 2

[0128] Example 2. VP1 truncated EV71 VLPs induce high levels of neutralizing antibodies

[0129] Truncated VLPs were massively expressed and purified from YE010 recombinant yeast. However, sufficient amount and purity of VLP-VP1Δ70 could not be purified from YE011 recombinant yeast for subsequent experiments. The protein composition of the truncated VLPs was determined by SDS-PAGE with reference to the full-length VLPs (designated VLP-FL). Such as image 3 As shown in A, compared with VLP-FL, VLP-VP1Δ27 has similar band patterns for VP0 and VP3, but different band patterns for VP1.

[0130] In order to study whether the truncation of VP1 protein affected the immunogenicity of VLPs, three groups (6 rats in each group) of ICR mice were immunized with VLP-FL, VLP-VP1Δ27 or PBS at week 0 and week 3 respectively, in which VLP- FL was the positive control group, and PBS was the negative control group. Serum samples were collected at week 5, and ELISA tests were performed to dete...

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
diameteraaaaaaaaaa
Login to View More

Abstract

The invention provides a novel virus-like particle derived from enterovirus 71 and its application. Specifically, after the VP1 protein of enterovirus 71 is truncated in the present invention, novel virus-like particles of enterovirus 71 can still be obtained, and the virus-like particles have high expression, strong immunogenicity, and good specificity.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular, the invention relates to a novel virus-like particle of enterovirus 71, its preparation method and application. Background technique [0002] Enterovirus 71 (Enterovirus 71, EV71) is a member of the Enterovirus genus of the Picornaviridae family, and is a non-enveloped single positive-sense RNA virus. The capsid of mature EV71 virus consists of 60 copies of VP1, VP2, VP3 and VP4. [0003] EV71 is the most important pathogen causing HFMD. Although EV71 infection usually manifests as a mild self-limited disease, it can also lead to severe neurological complications, such as brainstem encephalitis, poliomyelitis-like paralysis and pulmonary edema, and even death. As a candidate vaccine, EV71 virus-like particles (VLP) have been expressed in insect cell expression system and yeast expression system, and a large number of preclinical experiments have confirmed that it can induce neutralizin...

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 Patents(China)
IPC IPC(8): C12N7/04C07K14/085C12N15/41A61K39/125A61P31/14C12R1/93
Inventor 黄忠张超胡阳
Owner 中国科学院上海免疫与感染研究所
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com