Prefusion hPIV F Trimer Stabilization for Stronger Immune Response
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Solution Overview
Problem
Current vaccines for human parainfluenza viruses (hPIVs) are not available, and existing hPIV F ectodomain trimers do not effectively stabilize the prefusion conformation, leading to inadequate immune responses.
Innovation Solution
Development of recombinant hPIV1, hPIV2, hPIV3, and hPIV4 F ectodomain trimers with specific amino acid substitutions, deletions, and modifications that stabilize the prefusion conformation, including disulfide bonds, cavity-filling substitutions, and trimerization domains, which are incorporated into protein nanoparticles and administered with adjuvants.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing hPIV F ectodomain trimers are used without stabilization modifications, then the structure is simpler and easier to manufacture, but the immune response is inadequate
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid substitutions (such as cavity-filling substitutions at positions 466 and 473, and disulfide bond formations) to stabilize the F ectodomain trimer in its prefusion conformation. These molecular-level parameter changes enhance the structural stability and immunogenicity of the trimer, thereby improving the immune response effectiveness without fundamentally altering the overall trimer architecture
Solution Approach 2:
The patent employs composite material principles by incorporating multiple stabilization elements into the F ectodomain trimer structure, including disulfide bonds, cavity-filling amino acid substitutions, and trimerization domains. These composite structural features work synergistically to maintain the prefusion conformation and enhance immunogenicity, resolving the contradiction between structural simplicity and immune response effectiveness
2Reliability
If amino acid substitutions and modifications are introduced to stabilize prefusion conformation, then immune response is improved, but manufacturing complexity increases
Solution Approach 1:
The patent introduces specific amino acid substitutions (such as 162C and 168C disulfide bond formation, 170C and 242C disulfide bond, 213C and 230C disulfide bond, 216C and 221C disulfide bond, 85C and 222C disulfide bond, and 172C and 238C disulfide bond) to stabilize the prefusion conformation. These targeted parameter changes at the molecular level achieve conformational stability while maintaining compatibility with standard recombinant protein production methods
Solution Approach 2:
The patent applies preliminary action by designing the F ectodomain trimer with pre-installed stabilization features (disulfide bonds, cavity-filling substitutions, and trimerization domains) that are incorporated during the recombinant protein expression process. This preliminary structural stabilization ensures that the prefusion conformation is maintained throughout production and storage, reducing the need for complex post-manufacturing stabilization procedures
Data Source
AI summary
Embodiments of a recombinant human Parainfluenza Virus (hPIV) F ectodomain trimer stabilized in a prefusion conformation are provided. Also disclosed are nucleic acids encoding the hPIV F ectodomain trimer and methods of producing the hPIV F ectodomain trimer. Methods for inducing an immune response in a subject are also disclosed. In some embodiments, the method can be a method for treating or inhibiting a hPIV infection in a subject by administering a effective amount of the recombinant hPIV F ectodomain trimer to the subject.