Recombinant CSFV E2 Protein B/C Domain Swapping for DIVA Vaccines
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Solution Overview
Problem
Current CSFV vaccines lack the ability to differentiate between animals infected with the virus and those vaccinated, posing challenges in disease control and management.
Innovation Solution
A recombinant CSFV E2 protein is developed, where the 6B8 epitope is replaced by a corresponding fragment from a pestivirus, and additional mutations are introduced at specific amino acid positions to inhibit binding by the 6B8 monoclonal antibody, while maintaining immunogenicity and protective immunity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional CSFV vaccines are used, then protective immunity is achieved, but differentiation between infected and vaccinated animals is not possible
Solution Approach 1:
The patent applies local quality by introducing a specific marker epitope (6B8) at a defined location in the E2 protein structure. This localized modification allows the vaccine to maintain overall protective immunity while creating a specific distinguishable feature that enables differentiation between vaccinated and infected animals through targeted antibody detection.
Solution Approach 2:
The patent uses an intermediary approach by incorporating a foreign pestivirus epitope fragment into the CSFV E2 protein. This intermediary epitope serves as a marker that is recognized by specific antibodies, allowing indirect differentiation between vaccinated and infected animals without requiring direct detection of the virus itself.
2Loss of information
If the 6B8 epitope is replaced in the E2 protein, then differentiation capability is enabled, but antibody binding may be affected
Solution Approach 1:
The patent applies segmentation by dividing the E2 protein into functional domains and replacing only a specific epitopic region (amino acids 11-109) while preserving the rest of the protein structure. This segmented approach allows the marker function to be introduced without compromising the overall protein's ability to bind antibodies and elicit protective immunity.
Solution Approach 2:
The patent uses parameter changes by systematically modifying specific amino acid positions (10, 14, 22, 24, 25, 41, 64) in the E2 protein to create the 6B8 epitope variant. These controlled parameter changes in the protein sequence enable differentiation capability while maintaining sufficient antibody binding through conservation of other critical regions.
3Measurement precision
If multiple amino acid mutations are introduced, then epitope specificity is improved, but protein structure stability may be compromised
Solution Approach 1:
The patent applies segmentation by concentrating multiple amino acid mutations within a specific epitopic region (amino acids 11-109) rather than distributing them throughout the entire protein. This localized segmentation allows high epitope specificity to be achieved through multiple mutations while the rest of the protein structure remains intact and stable.
Solution Approach 2:
The patent uses local quality by introducing multiple amino acid changes (at positions 10, 14, 22, 24, 25, 41, 64) specifically within the epitope region to enhance specificity, while preserving the original amino acid sequence in other regions to maintain protein structure stability and folding characteristics.
Data Source
AI summary
The present invention relates the field of animal health. Particularly, the present invention relates to a recombinant classical swine fever virus E2 protein in which a fragment comprising at least one of the amino acids defining the 6B8 epitope of the CSFV E2 protein is replaced by a corresponding fragment of the E2 protein from a pestivirus other than CSFV. 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.