Unlock instant, AI-driven research and patent intelligence for your innovation.

Stabilized pre-fusion rsv f proteins

A pre-fusion, protein technology, applied in the medical field, can solve the problem of no vaccine and so on

Pending Publication Date: 2021-07-23
JANSSEN VACCINES & PREVENTION BV
View PDF24 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, efforts to produce vaccines against RSV have focused on developing vaccines containing prefusion forms of the RSV F protein (see, for example, WO20101149745, WO 2010 / 1149743, WO 2009 / 1079796, WO 2012 / 158613), but to date there is still no available vaccines

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
  • Stabilized pre-fusion rsv f proteins
  • Stabilized pre-fusion rsv f proteins
  • Stabilized pre-fusion rsv f proteins

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0127] Example 1 : Stability of prefusion conformations of RSV F variants in culture supernatants

[0128] The RSV F sequence used as a control for stability studies was based on the consensus sequence of the A subtype (SEQ ID NO: 13) or the consensus sequence of the B subtype (SEQ ID NO: 14), because the consensus sequence will correspond to the clinical isolate. The wild-type (non-passage) sequences of the strains are very similar (Kumaria et al. (2011)). For detection purposes, the F protein was fused at the C-terminus to a strep-tag. To assess the stability of various point mutations in RSV F, the amount of F protein in the prefusion conformation was measured in AlphaLISA.

[0129] By adding 5 μl of RSV F sample to 25 μl of prefusion specific Mab CR9501 (0.8 nM), anti-human IgG acceptor beads and streptavidin donor beads (Perkin Elmer) in The assay was performed in the mixture in assay buffer. After incubating the mixture for 2.5 hours at room temperature, the chemilu...

example 2

[0130] Example 2: Stable prefusion RSV F polypeptide - preparation of stabilizing mutations

[0131] Soluble prefusion proteins based on consensus RSV F (SEQ ID NO: 13 or 14) could not be purified due to apparent instability (e.g. image 3 , 4 shown). for evaluation image 3 Some of the stabilizing mutations shown in , in addition to the previously described D486N stabilizing mutation (Krarup et al., 2015), also produced two mutations containing a stable prefusion conformation, notably mutations P101Q and I152V in RSV-F protein (RSV180305; SEQ ID NO:20). In addition, another stable prefusion F protein (RSV172527; SEQ ID NO:21) was generated in which the P101Q and I152V substitutions were introduced in the prefusion RSV F protein, which already contained several previously described Stabilizing substitutions, in particular the mutations S46G, L203I, S215P, T357R, N371Y, D486N and D489Y. These constructs were synthesized and codon optimized at Gene Technologies, Inc. (Lif...

example 3

[0133] Example 3: SDS-PAGE analysis.

[0134] Purified RSV172527 was analyzed on 4%-12% (w / v) Bis-Tris NuPAGE gels, 1x MOPS (Life Technologies) under reducing or non-reducing conditions. All procedures were performed according to the manufacturer's instructions. For purity analysis, the gel was stained with Krypton Infrared protein stain (Thermo Scientific). Non-reduced and reduced RSV172527 were pure and in F 0 and F 1 Bands are separately visible at the expected height of the extracellular domain.

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 provides stable pre-fusion respiratory syncytial virus (RSV) F proteins, immunogenic compositions comprising said proteins and uses thereof for the prevention and / or treatment of RSV infection.

Description

[0001] The present invention relates to the field of medicine. The invention relates in particular to recombinant prefusion RSV F proteins, to nucleic acid molecules encoding these RSV F proteins, and to their use, eg in vaccines. Background technique [0002] After the discovery of respiratory syncytial virus (RSV) in the 1950s, the virus quickly became a recognized pathogen in humans associated with lower and upper respiratory tract infections. Worldwide, an estimated 64 million RSV infections occur annually, resulting in 160,000 deaths (WHO Acute Respiratory Infections updated September 2009). The most severe disease occurs especially in premature infants, the elderly, and immunocompromised individuals. In children under 2 years of age, RSV is the most common respiratory pathogen, accounting for approximately 50% of hospitalizations for respiratory infections, with the peak of hospitalizations occurring at 2–4 months of age. Almost all children have been reported to have ...

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): A61K39/12C07K14/005A61P31/14
CPCC07K14/005C07K2319/735A61K39/12A61P31/14C12N2760/18522C12N2760/18534
Inventor J·P·M·朗格戴克
Owner JANSSEN VACCINES & PREVENTION BV