Chimeric Polypeptides Using Six-Helix Bundles for Pre-Fusion Stability
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Solution Overview
Problem
Current vaccine designs for enveloped viruses and bacterial pathogens face challenges in stabilizing the pre-fusion conformation of viral fusion proteins, leading to premature conformational shifts and interference with HIV diagnostics, while lacking universal applicability and coverage for bacterial infections.
Innovation Solution
Development of chimeric polypeptides comprising microbial polypeptides connected to a heterologous structure-stabilizing moiety, specifically heptad repeat regions, forming a six-helix bundle to lock the pre-fusion conformation and elicit neutralizing antibody responses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If conventional recombinant expression of fusion proteins is used, then the protein can be produced, but it undergoes premature conformational shift to post-fusion form
Solution Approach 1:
A heterologous structure-stabilizing moiety is introduced as an intermediary element that binds to the fusion protein and stabilizes its pre-fusion conformation during recombinant expression. This mediator prevents premature conformational shifts while allowing the protein to be produced, resolving the contradiction between producibility and conformational stability.
Solution Approach 2:
The fusion protein is combined with a heterologous structure-stabilizing moiety to create a chimeric polypeptide. This composite structure leverages the stabilizing properties of the heterologous element to maintain the pre-fusion conformation, achieving both producibility and conformational stability.
2Stability of the object's composition
If HIV-based clamp is used for stabilization, then pre-fusion conformation is maintained, but diagnostic interference with HIV tests occurs
Solution Approach 1:
The essential stabilizing function of the HIV-based clamp is extracted and separated from the HIV-specific sequence elements that cause diagnostic interference. A heterologous structure-stabilizing moiety is used that provides the same stabilizing function without the harmful HIV-related immunogenicity, thereby eliminating diagnostic interference while maintaining pre-fusion conformation stability.
Solution Approach 2:
Instead of using the HIV-based clamp that causes long-term diagnostic interference, a heterologous stabilizing moiety is employed that provides temporary stabilization during expression and vaccination without causing persistent harmful effects. The heterologous element does not cross-react with HIV diagnostic tests, effectively replacing the problematic HIV-based solution.
3Stability of the object's composition
If target-specific structural vaccinology approaches are used, then pre-fusion stabilization is achieved, but universal applicability is limited
Solution Approach 1:
A universal heterologous structure-stabilizing moiety is developed that can be applied to multiple different fusion proteins from various viral and bacterial targets. This single stabilizing element provides pre-fusion conformation stability across different pathogens, eliminating the need for target-specific structural vaccinology approaches and enabling universal applicability while maintaining conformational stability.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The chimeric polypeptides effectively stabilize the pre-fusion conformation, enhancing immune responses against enveloped viruses and bacterial targets, reducing diagnostic interference, and providing a platform technology for diverse viral and bacterial infections.
Implementation Method 1
a heterologous structure-stabilizing moiety (SSM), wherein the structure-stabilizing moiety is a polypeptide comprising, in an N- to C-terminal order, a first heptad repeat region (FHRR) and second heptad repeat region (SHRR)... forming a six-helix bundle to lock the pre-fusion conformation
Data Source
AI summary
The present disclosure relates generally to chimeric polypeptides that comprise a microbial polypeptide (preferably (a) a virion surfaced exposed portion of an enveloped viral fusion protein or (b) a bacterial outer membrane polypeptide) and a heterologous structure-stabilizing moiety, and to complexes comprising those chimeric polypeptides. The present disclosure also relates to the use of these chimeric polypeptides and complexes thereof in compositions and methods for eliciting an immune response to a microbial polypeptide (preferably a fusion protein of an enveloped virus or a bacterial outer membrane polypeptide), or to respective complexes thereof and/or for treating or preventing related microbial infection (preferably an enveloped virus infection or a bacterial infection). Moreover, the disclosure further relates to compositions and methods for producing an antigen-binding molecule that specifically binds to such a microbial polypeptide or a complex thereof (preferably to an enveloped viral fusion protein or a complex thereof, or to a bacterial outer membrane polypeptide or a complex thereof).