Genetically engineered Foot and Mouth Disease Virus and related proteins, polynucleotides, compositions, methods and systems
Inventors
Rieder, Aida E. • De Los Santos, Teresa B. • Rodriguez, Luis L. • RAI, Devendra • DIAZ-SAN SEGUNDO, Fayna C. • Hoeprich, Paul D.
Assignees
University of Connecticut • Lawrence Livermore National Security LLC • US Department of Agriculture USDA
Publication Number
US-10953085-B2
Publication Date
2021-03-23
Expiration Date
2037-05-09
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Genetically engineered Foot and Mouth Disease Virus (FMDV) and related engineered proteins and polynucleotides, nanolipoprotein particles, compositions, methods and systems are described. The genetically engineered FMDV is modified by the strategic insertion of a protein tag into select regions of the FMDV genome which encode viral proteins that are exposed on the surface of the FMDV viral capsid. The inserted protein tag is displayed as a decoration or attachment on the viral capsid surface.
Core Innovation
The invention described is genetically engineered Foot and Mouth Disease Virus (FMDV) and related engineered proteins, polynucleotides, nanolipoprotein particles, compositions, methods and systems. The FMDV is modified by strategically inserting a protein tag into selected regions of the FMDV genome that encode viral proteins exposed on the viral capsid surface, leading to presentation of the protein tag on the external surface of the viral capsid.
The genetically modified FMD virus comprises a VP1 protein engineered to fuse at its C-terminus a 2A protein, with the protein tag inserted at specific positions relative to the N-terminus of the 2A protein or the C-terminus of the VP1 protein. This insertion allows the protein tag to decorate the viral capsid surface without compromising virus viability or antigenic properties.
The problem addressed arises from the difficulty in controlling Foot-and-Mouth Disease (FMD), a rapidly spreading viral disease in cloven-hoofed animals. Existing vaccines face challenges due to limited cross-protection between FMD strains of different serotypes and subtypes, and insufficient antigenic match within serotypes. Therefore, prevention and treatment of FMD remains challenging, necessitating improved vaccines and diagnostic tools.
Claims Coverage
The patent’s independent claims cover two main inventive features related to genetically modified FMD viruses comprising engineered VP1 proteins with inserted protein tags and nanolipoprotein particles functionalized to bind to the tagged viruses.
Genetically modified FMD virus comprising engineered VP1 protein with protein tag inserted at specific positions
A genetically modified Foot and Mouth Disease Virus (FMDV) having a viral capsid with an engineered VP1 protein that includes a protein tag of up to 100 amino acids inserted at an insertion position selected from any position −1 to +6 relative to the N-terminus of the 2A protein or from any position −1 to −7 relative to the C-terminus of the VP1 protein. The VP1 C-terminus is fused to the 2A N-terminus, enabling the external presentation of the protein tag on the viral capsid surface.
FMDV functionalized nanolipoprotein particle binding to tagged genetically engineered FMD virus
A functionalized nanolipoprotein particle that comprises a scaffold protein and a functionalized membrane forming lipid presenting a tag substrate configured to attach to the corresponding protein tag presented on one or more genetically engineered FMD viruses as described above. The tag substrate can be a molecule chelating a bivalent metal ion capable of specific binding to a polyhistidine tag on the virus, or other chemical groups specific to other protein tags.
The independent claims cover the genetic engineering of FMD viruses by insertion of protein tags at defined positions within the VP1 capsid protein fused to 2A protein, enabling surface display of tags for various applications, and also the design of nanolipoprotein particles functionalized with tag substrates that specifically bind these tagged viruses. These innovations allow improved purification, detection, vaccine formulation, and diagnostic uses of FMD viruses.
Stated Advantages
Provides viable genetically modified FMDV with tags presented on the viral capsid surface without affecting virus viability or entry into susceptible cells.
Enables specific binding of tagged FMDV to antibodies, divalent cations like Ni2+ and Co2+, and functionalized nanolipoprotein particles, allowing one-step purification and improved vaccine formulations.
Offers a marker vaccine permitting serological differentiation between vaccinated and infected animals (DIVA capability).
Facilitates rational design of vaccines incorporating engineered tags that interact with tag substrates and adjuvanted nanolipoprotein particles to improve immune response and protection.
Allows ease of exchange of capsid coding regions among different FMDV strains and serotypes via unique restriction sites, facilitating vaccine development against multiple strains.
Documented Applications
Use of genetically modified FMD viruses and tagged VP1 proteins for the production of vaccines capable of protecting cloven-hoofed animals against FMD.
Use of tagged FMD viruses for diagnostics, including differentiation of animals vaccinated with tagged virus versus naturally infected animals by serological assays.
Affinity purification of tagged FMD viruses using tag-substrate binding, such as polyhistidine tags binding to Ni2+ or Co2+ affinity columns or beads.
Formulation of vaccines comprising genetically modified FMD viruses with functionalized nanolipoprotein particles presenting corresponding tag substrates, optionally including adjuvants to enhance immune response.
Delivery and targeting of genetically modified FMD viruses to specific destinations using tag-mediated attachment to nanolipoprotein particles.
Use in research fields including basic and applied biology, bioengineering, medical diagnostics and therapeutics, vaccine development, and bioenergy.
Interested in licensing this patent?