Stabilized Pre-fusion RSV F Proteins via Amino Acid Mutations
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current vaccines based on the RSV F protein have failed due to stability, purity, and reproducibility issues, as they struggle to maintain the pre-fusion conformation of the RSV F protein, which is crucial for inducing effective neutralizing antibodies.
Innovation Solution
Development of stable, recombinant pre-fusion RSV F proteins stabilized in the pre-fusion conformation, using specific amino acid sequences and mutations, along with a heterologous trimerization domain, to create immunogenic compositions that induce neutralizing antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If conventional RSV F protein vaccine candidates are used, then they can potentially induce immune response, but they fail to maintain stable pre-fusion conformation leading to poor stability and reproducibility
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid substitutions (N67I, S215P, D386N) into the RSV F protein sequence to stabilize the pre-fusion conformation. These point mutations alter local structural parameters and energy landscapes, preventing unwanted conformational transitions to the post-fusion state, thereby resolving the contradiction between conformational stability and vaccine efficacy.
Solution Approach 2:
The patent creates a composite structure by fusing the RSV F protein with a heterologous stabilizing domain (alpha-helical coiled-coil domain from human fibroblast growth factor). This hybrid construct combines the immunogenic F protein with a structurally stable domain that templates and maintains the pre-fusion conformation, achieving both stability and immunogenicity simultaneously.
2Reliability
If pre-fusion conformation is targeted for vaccine design, then neutralizing antibody induction is improved, but protein stability and purity are compromised
Solution Approach 1:
The patent applies preliminary action by pre-stabilizing the F protein in the pre-fusion conformation through amino acid mutations and heterologous domain fusion before the protein is expressed and purified. The stabilizing elements are built into the protein structure in advance, ensuring that the protein folds into and remains in the desired pre-fusion conformation throughout production, purification, and storage, thereby maintaining both stability and neutralizing antibody induction capability.
Solution Approach 2:
The heterologous stabilizing domain acts as an intermediary that mediates the conformational stability of the F protein. This foreign domain serves as a structural template that binds to and stabilizes the pre-fusion conformation of the F protein without interfering with its immunogenic epitopes, thus resolving the contradiction between maintaining pre-fusion structure for antibody induction and ensuring overall protein stability.
3Reliability
If RSV F protein vaccine is developed, then protective immunity can be elicited, but manufacturing consistency and purity are difficult to achieve
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence parameters of the F protein (N67I, S215P, D386N substitutions) to create a version that is inherently more stable and less prone to conformational drift during manufacturing. These sequence changes make the protein folding landscape more robust, ensuring consistent production of the pre-fusion conformation across different batches and manufacturing conditions, thereby improving manufacturing precision while maintaining protective immunity.
Data Source
AI summary
Stable pre-fusion respiratory syncitial virus (RSV) F proteins (or fragment thereof) are described. Compositions containing the proteins and uses of the compositions for the prevention and/or treatment of RSV infection are also described.