Recombinant CSFV E2 Protein Epitope Mutation for Marker Vaccine Differentiation
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Solution Overview
Problem
Current CSFV vaccines lack differentiation between infected and vaccinated animals, leading to challenges in monitoring and controlling classical swine fever outbreaks, as existing vaccines do not provide a clear method to distinguish between animals infected with wild-type CSFV and those vaccinated with conventional vaccines.
Innovation Solution
A recombinant CSFV E2 protein with mutations in the 6B8 epitope is developed, allowing for the creation of an immunogenic composition that can differentiate between infected and vaccinated animals through specific immunological tests, utilizing a marker vaccine approach that inhibits the binding of the 6B8 monoclonal antibody, enabling effective monitoring and control of CSFV outbreaks.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional CSFV vaccines are used, then animals are protected from CSFV infection, but it becomes impossible to differentiate between infected and vaccinated animals
Solution Approach 1:
The patent applies local quality by introducing a specific mutation (R64W) at a localized epitope region (6B8 epitope) of the E2 protein. This localized change preserves the overall vaccine effectiveness while creating a detectable difference at a specific molecular location, enabling differentiation between vaccinated and infected animals through serological tests that target this specific epitope region.
Solution Approach 2:
The patent creates asymmetry by introducing a non-natural amino acid substitution (arginine to tryptophan at position 64) that breaks the symmetry of the epitope structure. This asymmetric change alters the epitope's recognition properties, allowing vaccinated animals to be distinguished from naturally infected animals whose viruses maintain the wild-type symmetric epitope structure.
2Loss of information
If a marker vaccine with epitope mutation is used, then differentiation between infected and vaccinated animals is enabled, but the vaccine must be carefully designed to maintain immunogenicity
Solution Approach 1:
The patent applies parameter changes by modifying a specific parameter (amino acid sequence) at a critical position (64) within the epitope region. This parameter change (R64W) is strategically selected to maintain the epitope's overall structure and immunogenicity while creating a detectable difference. The mutation is positioned to preserve T-cell epitope recognition and B-cell antibody binding, ensuring vaccine effectiveness is maintained.
Solution Approach 2:
The patent uses copying by creating a modified version of the wild-type E2 protein that replicates the overall immunogenic properties of the wild-type virus while incorporating a detectable marker. The recombinant E2 protein copies the essential features of the wild-type epitope (maintaining immunogenicity) while adding a specific mutation (R64W) that serves as a molecular fingerprint for differentiation.
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 recombinant CSFV E2 protein composition effectively prevents and treats CSFV diseases, provides a clear method to differentiate between infected and vaccinated animals, and aids in monitoring and controlling CSFV outbreaks by using a marker vaccine that inhibits the binding of the 6B8 monoclonal antibody, thereby enhancing public health and veterinary practices.
Implementation Method 1
A recombinant CSFV E2 protein with mutations in the 6B8 epitope is developed, allowing for the creation of an immunogenic composition that can differentiate between infected and vaccinated animals through specific immunological tests
Implementation Method 2
utilizing a marker vaccine approach that inhibits the binding of the 6B8 monoclonal antibody, enabling effective monitoring and control of CSFV outbreaks
Data Source
AI summary
The present invention relates to the field of animal health. Particularly, the present invention relates to a recombinant classical swine fever virus E2 protein comprising at least one mutation at the epitope specifically recognized by the 6B8 monoclonal antibody. Further, the present invention provides an immunogenic composition comprising the recombinant E2 protein of the present invention and the use of the immunogenic composition for preventing and/or treating diseases associated with CSFV in an animal. Moreover, the present invention provides a method and a kit for differentiating animals infected with CSFV from animals vaccinated with the immunogenic composition of the present invention. Furthermore, the present invention provides a method of producing the E2 protein.