Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Mutant of human papilloma virus 35 L1 protein

A protein and variant technology, applied in the fields of molecular virology and immunology, can solve problems such as safety issues and increased production costs of HPV vaccines

Active Publication Date: 2019-01-22
XIAMEN UNIV +1
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this approach will lead to a substantial increase in the production cost of the HPV vaccine, and may cause potential safety issues due to the increased dose of immunization

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
  • Mutant of human papilloma virus 35 L1 protein
  • Mutant of human papilloma virus 35 L1 protein
  • Mutant of human papilloma virus 35 L1 protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0175] Example 1. Expression and purification of mutated HPV35 L1 protein

[0176] Construction of expression vector

[0177] A multi-point mutation PCR reaction is used to construct an expression vector encoding an HPV35 L1 protein containing a mutation derived from segment 1 of the HPV16 L1 protein, wherein the initial template used is the pTO-T7-HPV35 L1 plasmid (which encodes the HPV35L1 protein; It is abbreviated as 35L1 in Table 2). The templates and primers used in each PCR reaction are shown in Table 2, and the amplification conditions of the PCR reaction are set as follows: denaturation at 94°C for 10 minutes; 7 minutes 30 seconds); a final extension of 10 minutes at 72°C. The specific sequences of the PCR primers used are listed in Table 3.

[0178] Add 2 μL DpnI restriction endonuclease to the amplification product (50 μL), and incubate at 37° C. for 60 min. 10 μL of the digested product was used to transform 40 μL of competent Escherichia coli ER2566 prepared...

Embodiment 2

[0210] Example 2: Assembly of HPV virus-like particles and detection of particle morphology

[0211] Assembly of HPV virus-like particles

[0212] Take a certain volume (about 2ml) of protein H35-16T1, H35-16T2, H35-16T3, H35-16T4, H35-16T5, H35-16T1-31S2, H35-16T1-31S3, H35-16T1-31S4, H35-16T1 -31S5, H35-16T4-31S1, H35-16T4-31S2, H35-16T4-31S3, H35-16T4-31S5 were respectively dialyzed into (1) 2L storage buffer (20mM sodium phosphate buffer pH 6.5, 0.5M NaCl ); (2) 2L refolding buffer (50mM sodium phosphate buffer pH 6.0, 2mM CaCl 2 , 2mM MgCl 2 , 0.5M NaCl); and (3) 20 mM sodium phosphate buffer pH 7.0, 0.5M NaCl. Dialysis was performed for 12 h in each of the three buffers.

[0213] By similar method, HPV35 L1, HPV16N30 and HPV31 L1 proteins were assembled into HPV35 VLP, HPV16N30 VLP and HPV31 VLP, respectively.

[0214] Molecular sieve chromatography analysis

[0215] A 1120 Compact LC high-performance liquid chromatography system from Agilent Corporation of the...

Embodiment 3

[0220] Example 3: Evaluation of thermal stability of virus-like particles

[0221]Use the differential temperature calorimeter VP Capillary DSC purchased from GE Company of the United States (formerly MicroCal Company) to evaluate H35-16T1, H35-16T2, H35-16T3, H35-16T4, H35-16T5, H35-16T1-31S2, H35- Thermal stability of VLPs formed by 16T1-31S3, H35-16T1-31S4, H35-16T1-31S5, H35-16T4-31S1, H35-16T4-31S2, H35-16T4-31S3, H35-16T4-31S5, wherein, The storage buffer of the protein was used as a control, and each protein was scanned from 10°C to 90°C at a heating rate of 1.5°C / min. Test results such as Figures 6A-6P shown. The results showed that the VLPs formed by each protein had extremely high thermal stability.

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
Radiusaaaaaaaaaa
Login to View More

Abstract

The invention relates to a mutant HPV35 L1 protein (or its variant), its coding sequence and a preparation method thereof. The invention also relates virus-like particles containing the protein. The protein (or its variant) and the virus-like particles are capable of inducing neutralizing antibodies against at least two types of HPV (such as HPV 35 and HPV 16, or HPV 35, HPV 16 and HPV 31), thereby preventing infections of the at least two types of HPV as well as diseases caused by the infections such as cervical cancer and condyloma acuminata. The invention also relates to application of theabove proteins and the virus-like particles to the preparation of a pharmaceutical composition or vaccine. The pharmaceutical composition or vaccine can be used for preventing the infections of the atleast two types of HPV as well as diseases caused by the infections such as cervical cancer and condyloma acuminata.

Description

technical field [0001] The present invention relates to the fields of molecular virology and immunology. Specifically, the present invention relates to a mutant HPV35 L1 protein (or its variant), its coding sequence and preparation method, and a virus-like particle comprising it, and the protein (or its variant) and the virus-like particle are capable of inducing Neutralizing antibodies against at least two types of HPV (e.g., HPV35 and HPV16, or HPV35, HPV16 and HPV31), thereby being useful for preventing infection by said at least two types of HPV and diseases resulting from said infection Such as cervical cancer and genital warts. The present invention also relates to the use of the above-mentioned protein and virus-like particles for preparing a pharmaceutical composition or a vaccine, which can be used to prevent the at least two types of HPV infection and the diseases caused by the infection Such as cervical cancer and genital warts. Background technique [0002] Hu...

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
IPC IPC(8): C07K14/025C12N15/37C12N7/00A61K39/12A61P31/20A61P35/00A61P17/12
CPCA61K39/12A61K2039/5258A61P17/12A61P31/20A61P35/00C07K14/005C12N7/00C12N2710/20022C12N2710/20023C12N2710/20034
Inventor 李少伟宋硕史晶洁何茂洲李智海夏宁邵
Owner XIAMEN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com