FMDV VLPs with Double Stabilizing Mutations
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Solution Overview
Problem
Current FMD vaccines based on inactivated virus are unstable, require high containment facilities, and have limited global availability, especially in tropical regions, due to their fragility and the need for a cold chain, which hinders effective immunization against foot-and-mouth disease.
Innovation Solution
Development of a recombinant foot-and-mouth disease virus (FMDV) VP2 protein with specific amino acid modifications (VP2-S093C and VP2-K190N) to enhance the stability and production yield of virus-like particles (VLPs), allowing for safer and more stable vaccine production in lower containment facilities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If inactivated FMD virus is used for vaccine production, then effective immunization against FMDV is achieved, but the vaccine becomes unstable and requires cold chain storage
Solution Approach 1:
The patent creates virus-like particles (VLPs) that are structural copies of the FMDV capsid but lack the viral genome. These VLPs replicate the immunogenic properties of intact FMDV capsids while eliminating the instability and safety concerns of using live or inactivated virus. The VLPs are produced by expressing the P1 capsid precursor protein in host cells, generating stable, non-infectious particles that maintain the conformational epitopes necessary for inducing protective immunity.
Solution Approach 2:
The patent extracts only the essential immunogenic component (the capsid structure) from the complete FMDV particle by removing the viral RNA genome. This extraction of the capsid building blocks (VP0, VP3, and VP1 proteins) that self-assemble into VLPs retains the protective antigenic properties while eliminating the harmful and unstable viral genetic material, thereby improving vaccine stability and safety.
2Reliability
If inactivated FMD virus is used for vaccine production, then effective immunization is achieved, but high containment facilities are required for production
Solution Approach 1:
The patent produces VLPs by expressing only the structural capsid proteins (P1 precursor) in host cells such as insect or mammalian cells, rather than cultivating complete FMDV particles. This copying approach using recombinant DNA technology eliminates the need for BSL-3 containment facilities, as the expression systems can be operated under lower biosafety levels while still generating the essential immunogenic capsid structures.
Solution Approach 2:
The patent separates the capsid protein coding sequence from the viral genome and expresses it independently in host cells. By extracting and independently expressing only the P1 capsid precursor gene, the production process eliminates the need to handle complete infectious virus particles, thereby reducing facility containment requirements from BSL-3 to BSL-2 or lower.
3Reliability
If inactivated FMD virus is used for vaccine production, then effective immunization is achieved, but production costs and facility maintenance costs are higher
Solution Approach 1:
The patent uses recombinant expression of VLPs in host cells (insect, mammalian, or plant systems) instead of traditional cell culture methods for growing FMDV. This copying approach using genetic engineering allows for scalable production in lower-containment facilities, reducing both capital investment and operational costs associated with maintaining high-containment infrastructure while maintaining vaccine effectiveness.
Solution Approach 2:
The patent employs mutagenesis and selection strategies to optimize the P1 capsid precursor protein for enhanced expression levels and improved stability of the resulting VLPs. By changing parameters such as codon optimization, protein stabilization mutations, and expression system conditions, the patent achieves higher yields of stable vaccine product, thereby reducing production costs and improving manufacturing efficiency.
4Stability of the object's composition
If VLP technology is used instead of inactivated virus, then product stability is improved, but production of stable VLPs requires optimized protein modifications
Solution Approach 1:
The patent introduces specific amino acid substitutions in the P1 capsid precursor protein sequence to enhance VLP stability. Examples include mutations such as S093C in VP2 that form stabilizing disulfide bonds, and other substitutions that strengthen inter-protein interactions within the capsid structure. These parameter changes in the protein sequence directly improve thermostability and pH resistance of VLPs without requiring complex production processes.
Solution Approach 2:
The patent creates VLPs with composite structural features by incorporating multiple stabilized protein components (VP0, VP3, VP1) that self-assemble into a robust capsid structure. The use of disulfide bonds and optimized protein-protein interfaces creates a composite-like structural integrity that enhances overall VLP stability, allowing the vaccine to withstand varying storage and transport conditions.
Data Source
AI summary
The invention concerns a modified recombinant foot and mouth disease virus (FMDV) VP2 protein and further concerns an FMDV capsid precursor protein P1 comprising the modified VP2 protein. In a specific aspect, the present invention concerns a VP2 protein or a capsid precursor protein P1 comprising the VP2 protein, wherein the amino acid sequence of the VP2 protein is modified to improve the stability of FMDV capsids. The invention further relates to an isolated nucleic acid molecule and an expression vector comprising the nucleic acid molecule for recombinant expression of the modified VP2 protein or a capsid precursor protein P1 comprising the VP2 protein. In further aspects, the invention relates to a virus-like particle (VLP) obtained from the modified capsid precursor protein P1 and a vaccine for use in the protection of a subject against an infection with FMDV produced from the VLP.