Stabilized HCMV Pentamer Complex via Polypeptide Mutations
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Solution Overview
Problem
Current HCMV vaccines face challenges in achieving effective immunogenicity due to the instability of the pentameric complex, which limits its preparation and production, and the presence of glycans that reduce the accessibility of neutralizing epitopes.
Innovation Solution
A stabilized HCMV pentamer complex is developed through recombinant expression, where at least one of the polypeptides is mutant, enhancing thermostability and epitope accessibility. This complex is used in an immunogenic composition or vaccine.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If the wild type pentameric complex is used for vaccine preparation, then the complex can be produced, but it exhibits poor thermostability which limits production and storage
Solution Approach 1:
The patent introduces mutations in the polypeptide sequences of the pentameric complex components (gH, gL, pUL128, pUL130, pUL131A) to change the physical and chemical parameters of the complex, specifically enhancing its thermostability. These mutations modify amino acid residues at specific positions to improve thermal stability without compromising the immunogenicity and neutralizing epitope integrity of the complex.
2Reliability
If glycans are present on the pentameric complex, then the complex is more native-like, but the glycans reduce accessibility of neutralizing epitopes to antibodies
Solution Approach 1:
The patent strategically removes or reduces glycans at specific positions on the pentameric complex where they would otherwise shield neutralizing epitopes from antibody access. By selectively eliminating these glycans while preserving the overall structure and other important regions, the invention enhances antibody binding to neutralizing epitopes without completely deglycosylating the complex, thus maintaining a balance between structural integrity and immunogenicity.
Data Source
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AI summary
Described are mutant, human cytomegalovirus (HCMV) pentamer complex polypeptides, methods of making them, and their use in HCMV protein complexes and compositions. In particular, the use of the modified HCMV polypeptides to stabilize HCMV complexes or unmask a pentamer epitope is described.