OMV Antigen Display via AMP Conjugation
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Solution Overview
Problem
Current OMV platforms face challenges in efficiently displaying a wide variety of antigens without requiring expression in the OMV-producing cell, limiting their versatility and immunogenicity, especially when using detergent extraction which can remove protective antigens and reduce stability.
Innovation Solution
A complex of Outer Membrane Vesicles (OMVs) with a non-covalently complexed vertebrate antimicrobial peptide (AMP) and an antigen, where the antigen is conjugated to the AMP, allowing for non-covalent attachment to the OMV surface, utilizing a detergent-free production method and genetic modifications to enhance OMV release and antigen presentation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If detergent extraction is used to increase OMV release, then OMV production efficiency is improved, but protective antigens are removed and long-term stability is reduced
Solution Approach 1:
The patent extracts and removes the detergent extraction step from the OMV production process. Instead of using detergents to increase OMV release, the invention uses a detergent-free approach where OMVs are harvested directly from culture supernatants, thereby preserving protective antigens and maintaining long-term stability while still achieving high yields through optimized culture conditions and genetic modifications for hypervesiculation
Solution Approach 2:
The patent changes the chemical parameters of the production system by eliminating detergents and instead using genetic modifications (such as mutations in lpxL, lpxA, lpxD, or rmpM genes) and optimized culture conditions to control OMV release. This parameter change allows high OMV production without the harmful effects of detergent extraction on antigen integrity and vesicle stability
2Adaptability or versatility
If endogenous expression is used for heterologous antigen display, then antigen presentation is improved, but the requirement for compatibility with bacterial outer membrane biogenesis machinery limits the variety of antigens that can be displayed
Solution Approach 1:
The patent segments the antigen display system into two independent components: (1) OMV production by the bacterial host, and (2) antigen production by a separate system (recombinant expression in other hosts or chemical synthesis). The antigens are then conjugated to OMVs through chemical coupling or physical association, eliminating the need for the antigen to be compatible with bacterial outer membrane biogenesis machinery and greatly expanding antigen versatility
Solution Approach 2:
The patent introduces an intermediary conjugation step that connects the antigen to the OMV surface. This intermediary approach allows antigens to be attached to OMVs without requiring them to be expressed by or integrated into the bacterial outer membrane machinery, thereby decoupling the complexity of antigen expression from the OMV production system and enabling display of diverse antigens including those from different pathogens
3Object-affected harmful factors
If LPS is genetically detoxified to optimize safety, then endotoxic activity is reduced, but adjuvant activity may be compromised
Solution Approach 1:
The patent applies precise parameter changes to the LPS structure through genetic modification, specifically targeting the lipid A region to reduce endotoxic activity while preserving adjuvant properties. By making targeted modifications (such as mutations in lpxL, lpxA, lpxD genes) rather than complete detoxification, the invention maintains the immunostimulatory capacity of LPS while reducing harmful endotoxic effects, achieving an optimal balance between safety and immunogenicity
Data Source
AI summary
The invention pertains to a complex of an OMV, a vertebrate antimicrobial peptide (AMP) and an antigen, wherein the AMP is non-covalently complexed with the OMV and wherein the antigen is conjugated to the AMP. Preferably, the antigen is covalently linked to the AMP. The invention further concerns the induction of an immune response using the complex of the invention as well as a method for producing the complex of the invention.