RSV F Protein Mutations for Stable Prefusion Trimer Antigens
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current vaccines targeting the prefusion conformation of the RSV F protein are unable to stabilize the protein in this state effectively, leading to a lack of an optimal antigen for inducing high neutralizing antibody titers.
Innovation Solution
Introduce mutations into specific regions of the RSV F protein, including the head and tail domains, to stabilize the prefusion trimer conformation, enhancing its stability and binding activity with monoclonal antibodies like AM14 and Synagis ®< .
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the RSV F protein is used in its natural prefusion conformation, then it can induce neutralizing antibodies, but it spontaneously converts to postfusion conformation and loses stability
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid mutations at positions 141, 190, 193, 195, 199, or 373 of the F protein sequence. These mutations alter the physical-chemical parameters of the protein structure, creating a stabilized prefusion conformation that resists spontaneous conversion to the postfusion state while maintaining immunogenicity.
Solution Approach 2:
The patent applies local quality by making targeted mutations at specific local regions (head domain positions 141, 190, 193, 195, 199 and position 373) rather than uniformly modifying the entire protein. This localized modification approach stabilizes the prefusion conformation at critical structural points while preserving the overall immunogenic properties of the F protein.
2Stability of the object's composition
If mutations are introduced to stabilize the prefusion conformation, then stability is improved, but the complexity of protein engineering increases
Solution Approach 1:
The patent applies segmentation by dividing the F protein into functional domains and identifying specific mutation sites within the head domain (positions 141, 190, 193, 195, 199) and position 373. This segmented approach allows systematic stabilization through targeted mutations at discrete locations, making the engineering process more manageable and predictable.
Data Source
Figure 1A~1B
Figure 2~3
Figure 4
AI summary
Disclosed is a mutant of a wild-type RSV F protein. The binding experiment of a pre-fusion conformation trimer-specific monoclonal antibody AM14 shows that the mutant provided in embodiments can have strong activity of binding to the AM14 monoclonal antibody, while the wild-type RSV F protein before artificial mutation has no activity of binding to the AM14 monoclonal antibody. The accelerated stability test result shows that the activity of binding of the mutant provided in the embodiments when placed in a 37°C environment for four weeks to the AM14 monoclonal antibody does not change obviously. The animal immune experiment result shows that compared with the wild-type RSV F protein before artificial mutation, the mutant provided in the embodiments can induce production of a neutralizing antibody having a higher titer.