SNARE-OMV Biotin Docking for Antigen Display
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Solution Overview
Problem
Current methods for loading foreign antigens onto outer membrane vesicles (OMVs) are hindered by challenges such as misfolding, proteolytic degradation, and inefficient bilayer translocation, especially for bulky and structurally complex antigens, leading to unpredictable expression and antigen density control, limiting the spectrum of cargo that can be packaged.
Innovation Solution
The development of a system using synthetic antigen receptors (SNAREs) comprising an outer membrane scaffold protein fused to a biotin-binding protein, allowing for the modular and controllable attachment of biotinylated antigens to the surface of OMVs through the AddVax platform, enabling the display of a diverse range of biomolecular cargo.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If foreign antigens are loaded onto outer membrane vesicles using current methods, then antigen display is achieved, but misfolding and proteolytic degradation occur leading to unpredictable expression
Solution Approach 1:
The patent introduces a biotin-binding protein as an intermediary component that mediates the attachment of biotinylated antigens to the outer membrane vesicle surface. This intermediary system avoids direct loading of antigens onto the OMV membrane, thereby preventing misfolding and proteolytic degradation while enabling predictable antigen expression through controlled biotin-biotinbinding protein interactions
Solution Approach 2:
The patent applies preliminary action by pre-modifying antigens with biotin groups before attachment to OMVs. This pre-biotinylation step ensures proper antigen folding and structural integrity prior to OMV association, preventing subsequent misfolding and degradation issues that occur with direct loading methods
2Adaptability or versatility
If current antigen loading methods are used, then some antigens can be displayed, but bulky and structurally complex antigens suffer from inefficient bilayer translocation
Solution Approach 1:
The biotin-binding protein serves as an intermediary that eliminates the need for bulky and structurally complex antigens to translocate through the OMV bilayer. Antigens are attached to the OMV surface via their biotin groups binding to the biotin-binding protein, allowing diverse cargo including large molecules to be displayed without requiring efficient bilayer translocation
Solution Approach 2:
The biotin-biotinbinding protein attachment system provides universal functionality for displaying diverse antigens on OMVs. This system can accommodate bulky and structurally complex antigens that cannot be efficiently translocated through the bilayer, thereby expanding the spectrum of packageable cargo while maintaining ease of manufacture through a unified attachment mechanism
3Manufacturing precision
If current methods are used for antigen loading, then antigen display is achieved, but antigen density control is unpredictable
Solution Approach 1:
The patent implements feedback control through the biotin-biotinbinding protein interaction system. By controlling the molar ratio of biotinylated antigen to biotin-binding protein and utilizing the high-affinity binding between them, the system provides predictable and controllable antigen density on OMV surfaces, with unbound biotin or biotin-binding protein serving as indicators of binding saturation
Solution Approach 2:
The patent applies parameter changes by controlling the biotinylation degree of antigens and the expression levels of biotin-binding protein on OMVs. By adjusting these parameters, precise control over antigen density is achieved, transforming the unpredictable expression into a controllable manufacturing parameter
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
This approach facilitates strong antigen-specific antibody responses and enables rapid vaccine development by allowing virtually any biotinylated antigen to be linked to OMVs, overcoming previous limitations in antigen display and expression.
Implementation Method 1
synthetic antigen receptor (SNARE) comprised of an outer membrane scaffold protein fused to a member of the avidin family... avidin-based dock-and-display for vaccine antigen cross (x)-linking
Data Source
AI summary
The present disclosure is directed to a system for displaying antigens. This system includes an outer membrane vesicle comprising a lipid bilayer and a synthetic antigen receptor comprising an outer membrane scaffold protein fused to a biotin-binding protein, where the outer membrane scaffold protein is incorporated in the lipid bilayer and the biotin-binding protein is displayed outside the outer membrane vesicle. Also disclosed are therapeutic compositions, nucleic acid constructs, expression vectors, and methods of eliciting an immune response in a subject.