Stabilizing HMPV F Protein Trimers for Vaccine Efficacy
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Solution Overview
Problem
Current efforts to develop an HMPV F protein-based vaccine have been unsuccessful due to challenges in stabilizing the HMPV F protein in its prefusion or postfusion conformation, which is crucial for inducing a robust immune response.
Innovation Solution
Recombinant HMPV F ectodomain trimers are engineered with specific amino acid substitutions to stabilize the protein in either the prefusion or postfusion conformation, including cysteine and proline substitutions to form non-native disulfide bonds and inhibit protease cleavage, enhancing stability and immunogenicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If the HMPV F protein is used in its native form, then it can be easily manufactured, but it cannot be stabilized in the prefusion or postfusion conformation required for effective vaccine function
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid substitutions (cysteine and proline substitutions) at defined positions in the HMPV F protein sequence. These substitutions alter the chemical parameters of the protein to form non-native disulfide bonds that lock the protein in stable prefusion or postfusion conformations, directly resolving the conformational instability problem while maintaining a relatively simple implementation approach
Solution Approach 2:
The patent creates composite protein structures by combining the HMPV F protein with engineered disulfide bond networks. The introduction of non-native disulfide bonds through cysteine substitutions creates a composite structural system where the native protein framework is reinforced with artificial cross-links, achieving conformational stability without significantly increasing overall structural complexity
2Stability of the object's composition
If amino acid substitutions are introduced to stabilize the F protein conformation, then conformational stability is improved, but the manufacturing process becomes more complex
Solution Approach 1:
The patent modifies the amino acid sequence parameters by introducing specific cysteine and proline substitutions at predetermined positions. These parameter changes enable the formation of stabilizing disulfide bonds and structural constraints that lock the protein in desired conformations, achieving stability while maintaining straightforward manufacturing through standardized site-directed mutagenesis techniques
3Stability of the object's composition
If the F protein is proteolytically processed to generate F1 and F2 polypeptides, then the native processing pathway is followed, but the prefusion conformation cannot be maintained for vaccine purposes
Solution Approach 1:
The patent applies preliminary anti-action by introducing proline substitutions at strategic positions that prevent proteolytic cleavage of the F protein. These substitutions create steric or structural barriers that block protease access to cleavage sites, thereby preventing the native processing pathway that would otherwise destroy the prefusion conformation, while maintaining the protein's immunogenic functionality
Solution Approach 2:
The patent changes the chemical parameters of the F protein by introducing proline substitutions that alter the local structure and resistance to proteolysis. These parameter changes increase the protein's reliability by preventing unwanted cleavage, ensuring the protein maintains its intended conformation and functional properties throughout storage and administration
Data Source
AI summary
Metapneumovirus (HMPV) F ectodomain trimers stabilized in a prefusion or postfusion conformation, nucleic acid molecules and vectors encoding these proteins, and methods of their use and production are disclosed. In several embodiments, the HMPV F ectodomain trimers and/or nucleic acid molecules can be used to generate an immune response to HMPV in a subject. In additional embodiments, the therapeutically effective amount of the HMPV F ectodomain trimers and/or nucleic acid molecules can be administered to a subject in a method of treating or preventing HMPV infection.