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

Influenza virus hemagglutinin mutants

An influenza virus, hemagglutinin technology, applied in the direction of viruses/phages, viruses, viral peptides, etc., can solve problems such as low efficacy

Pending Publication Date: 2021-04-09
MEDICAGO INC
View PDF38 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mutant hemagglutinin protein maintained potency at 25°C for at least 12 months, while wild-type hemagglutinin protein exhibited less than 40% potency after only 50 days after purification

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
  • Influenza virus hemagglutinin mutants
  • Influenza virus hemagglutinin mutants
  • Influenza virus hemagglutinin mutants

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0308] Example 1: Influenza virus hemagglutinin constructs

[0309] Influenza virus hemagglutinin constructs are generated using techniques well known in the art. For example, wild type H3 A-Switzerland / 9715293 / 13 HA, N382A A / Switzerland / 9715293 / 13 H3 hemagglutinin and CysTM A-Switzerland-9715293-13 were cloned as described below. Other H3 hemagglutinin mutants were obtained using similar techniques, and the hemagglutinin sequence primers, templates and products are illustrated in Example 3 (Production of influenza virus hemagglutinin and VLPs in plants) and Table 4.

[0310] Table 4 below summarizes wild-type and mutant hemagglutinin proteins, primers, templates and products. For the H3 influenza virus constructs other than the A / Switzerland / 9715293 / 13 hemagglutinin protein cloned into the 1190 cloning vector without M2, the cloning vector used incorporated the M2 influenza virus ion channel gene, the M2 influenza Viral ion channel genes are under the control of the alfal...

example 2

[0318] Example 2: Method

[0319] Agrobacterium tumefaciens transfection

[0320] Agrobacterium tumefaciens strain AGL1 was transfected by electroporation with wild-type influenza virus hemagglutinin or mutant influenza virus hemagglutinin expression vector using the method described by D'Aoust et al. (Plant Biotech.J.6:930-40) . Make the transfected Agrobacterium in the addition of 10 mmol 2-(N-morpholino)ethanesulfonic acid (MES), 20 μM acetosyringone, 50 μg / ml kanamycin and 25 μg / ml carbenicillin, Grow in YEB medium at pH 5.6 until the OD600 reaches between 0.6 and 1.6. The Agrobacterium suspension was centrifuged and resuspended in osmotic medium (10 mmol MgCl2 and 10 mmol MES, pH 5.6) before use.

[0321] Plant biomass, preparation of inoculum, and Agroinfiltration

[0322] Nicotiana benthamiana plants were grown from seed in flats filled with a commercially available peat moss substrate. The plants were grown in a greenhouse under a 16 / 8 photoperiod and a temp...

example 3

[0332] Example 3: Production of influenza virus hemagglutinin protein in plants

[0333] Modification of H3 hemagglutinin I

[0334] Influenza virus hemagglutinin constructs were generated using techniques well known in the art (see Example 1). Table 4 below summarizes wild-type and mutant hemagglutinin proteins, primers, templates and products. The sequences used are listed in Example 4 and in the Sequence Listing.

[0335] N382A A / Switzerland / 9715293 / 13 mutant H3

[0336] N382A A / Switzerland / 9715293 / 13 mutant H3 (construct number 3023) was constructed by mutating the asparagine at position 382 of wild-type / Switzerland / 9715293 / 13 H3 to alanine. like figure 2As shown, the purified extract of N. benthamiana plants infiltrated with construct No. 3023 exhibited approximately 30% increased hemagglutination titer.

[0337] L384V A / Switzerland / 9715293 / 13 mutant H3

[0338] L384V A / Switzerland / 9715293 / 13 mutant H3 (construct number 3034) was constructed by mutating leucine ...

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

No PUM Login to View More

Abstract

The present invention relates to the production of modified influenza viral proteins in plants. More specifically, the present invention relates to producing and increasing influenza virus-like particle (VLP) production in plants, wherein the VLPs comprise the modified influenza viral proteins, such as modified influenza hemagglutinin (HA). The HA protein may comprising an amino acid sequence comprising at least one substitution when compared to a corresponding wildtype amino acid sequence. Further provided are nucleic acid encoding the modified HA protein. Furthermore methods of producing an influenza virus like particle (VLP) and methods of increasing yield of production of an influenza virus like particle (VLP) in a plant, portion of a plant, or a plant cell, are also provided.

Description

technical field [0001] The present invention relates to the production of mutant viral proteins in plants. More specifically, the present invention relates to the production of influenza virus-like particles and increased production of influenza virus-like particles in plants. Background technique [0002] Influenza viruses are enveloped single-stranded RNA viruses of the Orthomyxoviridae family. Influenza viruses are highly contagious and can cause mild to severe illness in all age groups. [0003] Vaccination remains the most effective way to prevent influenza infection. Vaccinations are traditionally done using live attenuated or completely inactivated viruses, which elicit an immune response when administered to a patient. To eliminate the potential risk of both live attenuated and fully inactivated viruses regaining the ability to replicate and infect, vaccines including recombinant viral proteins are also used to elicit protective immunity against influenza infectio...

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): C12N15/44C07K14/11C12N15/82C12N15/84C12N5/10A01H5/00A01H6/82C07K16/10A61K39/145A61P31/16A61P37/04
CPCC07K14/11C07K16/10A61P37/04A61P31/16C12N15/8258A61K2039/5258A61K39/12C12N2760/16134C12P21/02C12N7/00C12N2760/16123C07K14/005A61K39/145C07K16/1018A61K38/00C12N2760/16122C12N2760/16152C12N2760/16023C12N2760/16033C12N15/8257
Inventor 皮尔-奥利弗·拉沃伊奥雷连·洛瑞阿兰·道赛特马克-安德鲁·德奥斯特玛南·科沃托
Owner MEDICAGO INC
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