Bordetella OmpA D124N Mutation for Higher OMV Production
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Solution Overview
Problem
There is a need to increase OMV production in Bordetella bacteria for improved vaccine development, particularly for Bordetella pertussis, as current methods do not optimize secretion rates of spontaneously formed OMVs, and there is a requirement for a modified Bordetella with increased immunogenicity and blebbing phenotype.
Innovation Solution
A mutation in the OmpA-like domain, specifically a D124N substitution at position 124, enhances OMV production in Bordetella bacteria, including strains such as B. pertussis, B. parapertussis, and B. bronchiseptica, leading to a high-blebbing phenotype and increased immunogenicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If wild-type Bordetella bacteria are used, then natural OMV production occurs, but the secretion rate is insufficient for optimized vaccine production
Solution Approach 1:
The patent applies parameter changes by modifying the OmpA protein sequence through site-directed mutagenesis. Specifically, amino acid substitutions (such as D124N, D124E, or D124K) are introduced at position 124 of the OmpA protein to alter its properties and enhance OMV secretion. This targeted parameter change in the protein structure directly increases both the secretion rate and production amount of OMVs, resolving the contradiction between natural production limitations and vaccine production requirements.
2Reliability
If aCPS vaccines are used, then pertussis protection is provided, but immunogenicity and protective efficacy are insufficient
Solution Approach 1:
The patent utilizes parameter changes by modifying the OmpA protein sequence to enhance the immunogenic properties of OMVs. The amino acid substitutions in OmpA not only increase OMV secretion but also improve the quality and immunogenicity of the vesicles. This dual effect addresses the contradiction by simultaneously enhancing both the quantity of OMVs produced and their protective efficacy, making them suitable for improved acellular pertussis vaccines.
Data Source
AI summary
The invention pertains to a mutant Bordetella OmpA polypeptide. A Bordetella comprising said mutant polypeptide has a high blebbing phenotype. Hence, the invention further pertains a method for producing OMVs, wherein the method comprises a step of culturing a population of Bordetella bacteria comprising a modified OmpA polypeptide, under conditions conductive for the production of OMVs. In addition, the invention pertains to OMVs produced by a Bordetella comprising a mutant OmpA polypeptide, and the use of such OMVs for the treatment and prevention of a Bordetella infection.