Antigen-Presenting Vesicles Using Biotin-Avidin Antigen Coupling
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Solution Overview
Problem
Existing conjugation technologies for antigen coupling in antigen-presenting vesicles (APV) risk denaturation and structural damage to proteins and vesicles, leading to ineffective immunogenic compositions.
Innovation Solution
A process involving biotinylation of outer membrane vesicles (OMVs) using biotin or derivatives, combined with recombinant avidin-like molecules, allows controlled conjugation of antigens without chemical interference, preserving antigenicity and vesicle structure.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If typical conjugation technology is used to couple antigens to vesicles, then antigen coupling is achieved, but the risk of denaturation and structural damage to the antigen protein increases
Solution Approach 1:
The patent introduces biotin as an intermediary molecule that mediates the coupling between antigens and vesicles. The biotin-avidin interaction serves as a gentle, non-denaturing bridge that allows antigen attachment without direct harsh chemical conjugation to the antigen itself, thereby preserving antigen protein stability while achieving coupling
Solution Approach 2:
The patent changes the chemical parameters of the conjugation process by using biotinylation chemistry that targets specific functional groups (carboxylic groups) on the vesicle surface rather than using rigorous conjugation conditions. This parameter change allows coupling at milder conditions that preserve protein integrity
2Manufacturing precision
If rigorous conjugation treatment is applied to couple antigens, then coupling efficiency is improved, but unnecessary modifications and damage to the vesicle structure occur
Solution Approach 1:
The patent applies local quality by targeting specific carboxylic functional groups on the vesicle surface for biotinylation rather than applying rigorous conjugation throughout the entire vesicle structure. This localized modification approach achieves efficient antigen coupling while leaving the rest of the vesicle structure intact and undamaged
Solution Approach 2:
Biotin serves as a localized intermediary that concentrates the coupling function at specific sites on the vesicle surface, allowing efficient antigen attachment without requiring global structural modification or damage to the vesicle
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 process ensures effective coupling of antigens to OMVs, maintaining their immunogenicity and structural integrity, enabling the production of stable antigen-presenting vesicles for vaccines and immunotherapeutics.
Implementation Method 1
EDAC, activator molecule, reacts with the carboxylic groups, activating them and making them available for conjugation with the amine groups of the biotin derivative
Implementation Method 2
at least a pair of molecules with complementary affinities (for example, a biotin or a derivative and a protein with affinity to biotin)
Data Source
Figure 1(A)~2(B)
Figure 3(A)~3(D)
Figure 4
AI summary
The present invention refers to a process for obtaining antigen-presenting vesicles (APV) that enables the coupling of one or more antigens, wherein such APV comprises (i) an outer membrane vesicle of gram-negative bacteria (OMV); (ii) at least one antigenic protein or peptide; and (iii) at least a pair of molecules with complementary affinities comprising a first affinity molecule that associates with the vesicle, and a complementary affinity molecule that associates with the protein or peptide. Thus, the process for obtaining the APV of the present invention is essential to achieve a form of presentation that consists of a vesicle that makes it possible to attach one or more proteins, or a plurality of different protein or peptide antigens. In that regard, such process comprises the following steps: (a) conjugating the first affinity molecule to the vesicle (OMV); (b) obtaining the antigen protein(s) or peptide(s) in fusion with the complementary affinity molecule; and (c) coupling the fusion protein obtained in step "b" with the product obtained in step "a".