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

Rsv f protein compositions AMD methods for making same

a technology of protein composition and rsv, which is applied in the field of rsv f protein composition, can solve the problems of purification, homogeneous, and difficult to obtain immunogenic preparations, structure and refolding,

Inactive Publication Date: 2012-06-28
NOVARTIS AG
View PDF2 Cites 28 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes immunogenic compositions that contain RSV F polypeptides. These polypeptides can be soluble or in the form of a trimer. The RSV F polypeptides can have mutations that improve their function in the body. The immunogenic compositions can also contain an adjuvant to enhance their immunogenicity. The technical effect of the patent is to provide improved immunogenic compositions for the prevention and treatment of RSV infections.

Problems solved by technology

Due to the complexity of RSV F protein processing, structure and refolding, purified, homogeneous, immunogenic preparations are difficult to obtain.

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
  • Rsv f protein compositions AMD methods for making same
  • Rsv f protein compositions AMD methods for making same
  • Rsv f protein compositions AMD methods for making same

Examples

Experimental program
Comparison scheme
Effect test

example 1

RSV F Polypeptides

[0467]This example provides sequences of a number of examples of polypeptides (e.g., that contain signal sequences) and nucleic acid sequences that may be used to express RSV F polypeptides of the present invention. The presented amino acid sequences include the signal peptide and contain an optional C-terminal linker and His tag (GGSAGSGHHHHHH (SEQ ID NO:90)). When these polypeptides are produced in host cells, the polypeptide will usually be processed by the cell to remove the signal peptide and, as described herein, some of the polypeptides will be cleaved, for example at unmodified furin cleavage sites. The invention includes compositions that contain, all forms of the particular RSV F protein ecto-domain polypeptides disclosed herein, including mature forms, which lack the signal peptide, forms that may be cleaved into subunits that comprise F1 and F2, and forms that lack the optional C-terminal His tag. The following examples are merely illustrative of the sc...

example 2

Expression and Purification of RSV F Constructs

[0486]The RSV F ECTO and truncated constructs, lacking the transmembrane domain and cytoplasmic tail region with either wild-type furin cleavage sites or harboring knock-out mutations to the furin cleavage sites and with or without prefusion stabilization mutations, were cloned into a pFastBac baculovirus expression vector (Invitrogen). Several of these constructs contain a C-terminal flexible linker followed by a His6-tag sequence used for chelating purification. The production of high-titer baculovirus stocks were passaged in Sf9 insect cells. Proteins were expressed by infecting either Sf9, Tn5 or High Five insect cells with the required baculovirus and harvesting the media supernatant two or three days post infection, monitored by western blot using an anti-RSV F or anti-6HIS antibody.

[0487]Large scale expression media was concentrated / purified by one of two general strategies for eliminating the deleterious Effect of the ferritin p...

example 3

Detection of Pre-Fusion and Post-Fusion RSV F

[0496]A number of methods are available to determine the conformation of the RSV F protein to assay whether a modification to the RSV F polypeptide or added molecule disfavors the post-fusion conformation. Examples include liposome association, conformation specific monoclonal antibodies (including as used in FACS, ELISA, etc.), electron microscopy, differential protease sensitivity between the conformations, gel filtration chromatography, analytical ultracentrifugation, dynamic light scattering, deuterium exchange NMR experiments, mass spectroscopy, circular dichroism spectroscopy, isothermal titration calorimetry, tryptophan spectroscopy, and X-ray crystallography.

Liposome Association

[0497]Liposome association may be used to assay the conformation of the RSV F protein. Soluble forms of the RSV F protein in the pre-fusion conformation will not associate with liposomes while the post-fusion conformation will associate with liposomes.

[0498...

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
immunogenic compositionaaaaaaaaaa
Login to View More

Abstract

The present invention relates to immunogenic compositions comprising RSV F protein, methods for preparing compositions that contain RSV F protein ecto-domain polypeptides, and to certain engineered RSV F proteins and nucleic acids that encode the engineered RSV F proteins. Compositions prepared using the methods can contain RSV F protein ecto-domain polypeptides in a predominant or single desired form and conformation. The invention also relates to methods for inducing an immune response to RSV F.

Description

RELATED APPLICATIONS[0001]This application claims the benefit of U.S. Patent Application No. 61 / 225,805, filed on Jul. 15, 2009, and U.S. Patent Application No. 61 / 294,426, filed on Jan. 12, 2010. The entire teachings of the above applications are incorporated herein by reference.BACKGROUND OF THE INVENTION[0002]Respiratory syncytial virus (RSV) is an enveloped non-segmented negative-strand RNA virus in the family Paramyxoviridae, genus Pneumovirus. It is the most common cause of bronchiolitis and pneumonia among children in their first year of life. RSV also causes repeated infections including severe lower respiratory tract disease, which may occur at any age, especially among the elderly or those with compromised cardiac, pulmonary, or immune systems.[0003]To infect a host cell, paramyxoviruses such as RSV, like other enveloped viruses such as influenza virus and HIV, require fusion of the viral membrane with a host cell's membrane. For RSV the conserved fusion protein (RSV F) fu...

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(United States)
IPC IPC(8): A61K39/155A61P31/14C07K14/135
CPCA61K39/00C07K2317/76A61K2039/5258A61K2039/53A61K2039/55505A61K2039/55555A61K2039/55561A61K2039/55572C07K14/005C07K16/1027C07K2319/00C07K2319/21C07K2319/50C07K2319/73C12N2760/18522C12N2760/18534A61K2039/5252A61K2039/5256A61K2039/55566C12N2770/36143A61K39/155A61K39/12A61P11/00A61P31/14A61P37/04
Inventor SWANSON, KURTDORMITZER, PHILIP
Owner NOVARTIS AG
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