Stabilized Pre-fusion RSV F Polypeptides for Vaccine Immunogenicity
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current vaccine candidates based on the RSV F protein have failed due to stability, purity, and reproducibility issues, particularly in maintaining the pre-fusion conformation, which is essential for eliciting protective neutralizing antibodies.
Innovation Solution
Development of stable, recombinant pre-fusion RSV F polypeptides stabilized in the pre-fusion conformation, using mutations and trimerization domains like the GCN4-pII or fibritin trimerization domain, to create soluble and stable immunogenic compositions that induce neutralizing antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If vaccine candidates are based on RSV F protein to elicit neutralizing antibodies, then immunogenicity is improved, but stability and conformational maintenance deteriorate
Solution Approach 1:
The RSV F protein is segmented into F1 and F2 domains with specific truncations (e.g., F1 domain truncated at amino acid 495 or 513) to create stable pre-fusion conformations while retaining immunogenic epitopes. This segmentation allows the protein to maintain its protective conformation without undergoing complete refolding to post-fusion state.
Solution Approach 2:
Amino acid substitutions are introduced at specific positions (e.g., Q491, Q492, Q493, Q494 in F1 domain; K513, K514, K515, K516 in F2 domain) to stabilize the pre-fusion conformation. These parameter changes in the protein sequence prevent unwanted conformational changes while preserving the ability to elicit neutralizing antibodies.
2Reliability
If RSV F protein is used as vaccine candidate, then neutralizing antibody induction is improved, but purity and reproducibility deteriorate
Solution Approach 1:
The transmembrane domain and cytoplasmic tail are removed from the full-length RSV F protein, extracting only the extracellular domain (ECD) that contains the immunogenic epitopes. This extraction creates a soluble protein that is easier to purify and produces more consistent results across manufacturing batches.
Solution Approach 2:
The protein is engineered with stable pre-fusion conformation through domain truncation and amino acid substitutions before vaccination. This preliminary stabilization ensures that the protein maintains its protective conformation throughout storage and administration, guaranteeing consistent immunogenicity across different batches.
3Reliability
If pre-fusion conformation is maintained for vaccine efficacy, then neutralizing antibody response is improved, but protein stability deteriorates
Solution Approach 1:
The RSV F protein is combined with stabilizing elements including specific amino acid substitutions and domain truncations to create a composite structure that locks the pre-fusion conformation. This composite approach integrates multiple stabilizing features into a single protein construct that maintains both stability and immunogenicity.
Solution Approach 2:
Amino acid substitutions are introduced at positions that prevent the conformational change from pre-fusion to post-fusion state. These preliminary anti-actions counteract the natural tendency of the protein to refold, effectively blocking the transition and stabilizing the protective pre-fusion conformation for long-term storage.
Data Source
AI summary
Described are stable pre-fusion respiratory syncitial virus (RSV) F polypeptides, immunogenic compositions comprising the polypeptides and uses thereof for the prevention and/or treatment of RSV infection.