Modified FMDV 3C Protease Reduces Host Cell Cytotoxicity
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Solution Overview
Problem
Current foot-and-mouth disease virus (FMDV) vaccines face limitations due to the cytotoxicity of native FMDV 3C protease, which impairs the expression of immunogenic antigens in host cells, leading to poor recombinant yields and stability issues, and existing vaccines have short shelf life and induce insufficient immunity.
Innovation Solution
Engineering a modified FMDV 3C protease with specific amino acid substitutions, such as L127P, to reduce cytotoxicity while maintaining proteolytic activity, allowing for efficient processing of FMDV precursor polypeptides into immunogenic proteins without significant host cell toxicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If native FMDV 3C protease is used to process FMDV precursor polypeptides, then viral proteins are produced, but host cell cytotoxicity increases and recombinant yields decrease
Solution Approach 1:
The patent applies parameter changes by mutating specific amino acid residues in the FMDV 3C protease sequence (e.g., positions 46, 84, 163, and others) to alter the enzyme's properties. These mutations modify the protease's specificity and reduce its cytotoxic effects on host cells while maintaining its ability to process viral precursor polypeptides, thereby improving recombinant protein yields.
Solution Approach 2:
The invention applies local quality by making specific localized mutations at particular amino acid positions within the protease molecule (such as the catalytic triad residues and other specific positions) rather than uniformly modifying the entire protein. This allows the protease to maintain its essential function in specific regions while reducing harmful cytotoxic properties in other regions.
2Adaptability or versatility
If whole virus vaccines are used, then broad spectrum immunity is provided, but vaccine stability and shelf life are poor
Solution Approach 1:
The patent applies segmentation by dividing the whole virus vaccine into separate components: recombinant viral antigens produced in host cells and modified FMDV 3C protease. This segmentation allows the antigens to be produced without the instability issues of whole virus formulations, while the modified protease with reduced cytotoxicity enables better production yields and stability.
Solution Approach 2:
The invention applies copying by using recombinant DNA technology to produce viral antigens in host cells rather than using whole virus particles. The modified FMDV 3C protease facilitates this copying process by efficiently processing viral precursor polypeptides into immunogenic antigens with reduced harm to host cells, thereby improving vaccine stability and shelf life while maintaining broad spectrum immunity.
3Quantity of substance
If FMDV 3C protease is expressed in host cells to produce viral antigens, then antigen production increases, but host cell viability decreases
Solution Approach 1:
The patent applies parameter changes by mutating specific amino acid residues in the FMDV 3C protease (including positions 46, 84, 163, and others) to alter the enzyme's properties. These mutations reduce the protease's cytotoxic effects on host cells while maintaining its ability to process viral precursor polypeptides, thereby improving both antigen production and host cell viability.
Solution Approach 2:
The invention applies 'blessing in disguise' by converting the harmful cytotoxic property of FMDV 3C protease into a beneficial trait through targeted mutations. The modified protease retains its essential function of processing viral antigens while its reduced cytotoxicity becomes a benefit, allowing host cells to remain viable and continue producing antigens at high levels.
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 modified FMDV 3C protease effectively processes FMDV P1 precursor into viral proteins, enhancing recombinant yields and stability, and increases the production of immunogenic proteins, leading to improved vaccine efficacy and longer shelf life.
Implementation Method 1
the modified FMDV 3C protease effectively processes FMDV P1 precursor into viral proteins
Data Source
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AI summary
This application is directed generally to foot-and-mouth disease virus (FMDV) 3C proteases that have, been modified by mutating a polynucleotide sequence coding for the FMDV 3C protease. The modified FMDV proteases exhibit proteolyic activity on FMDV Fl precursor protein and exhibit a reduction in one or more toxic or inhibitory properties associated with an unmodified FMDV 3C protease on a host cell used to recombinantly produce it. Vectors carrying polynucleotides encoding modified FMDV 3C protease sequences can induce production of FMDV virus-like particles in a host cell when expressed in the host cell. The modified FMDV 3C proteases can generally be used to produce immunogenic FMDV preparations capable of inducing an immune response against FMDV.