The invention features modified alphavirus or flavivirus virus-like particles (VLPs). The invention provides methods, compositions, and kits featuring the modified VLPs. The invention also features methods for enhancing production of modified VLPs for use in the prevention or treatment of alphavirus and flavivirus-mediated diseases. The invention also provides methods for delivering agents to a cell using the modified VLPs.

The present invention provides compositions and methods for therapeutic immunization for treatment of infectious diseases and/or cancer. Methods of the invention include a method generating a high titer infectious agent and cancer vector, methods of treating and/or preventing cancer or an infection by an infectious disease, and methods of inducing a memory T and B cell immune response against infectious agent and cancer in a subject administered the VLV composition produced thereby. Furthermore, the invention encompasses a pharmaceutical composition for vaccinating a subject to protect the subject against cancer or an infectious agent.

Described herein are i-DNA vectors and vaccines and methods for using the same. The i-DNA generates live attenuated vaccines in eukaryotic cells in vitro or in vivo for pathogenic RNA viruses, particularly chikungunya virus (CHIKV). When iDNA is injected into the vaccine recipient, RNA of live attenuated virus is generated by in vivo transcription in the recipient's tissues. This initiates production of progeny attenuated viruses in the tissues of the vaccine recipient, as well as elicitation of an effective immune response protecting against wild-type, non-attenuated virus.

The invention provides a cell-free system comprising not more than about 5% wheat germ extract for expressing proteins such as viral proteins and proteins required for viral capsid assembly, and proteins that assemble into multiprotein complexes in a manner analogous to viral capsids, are provided. Further provided are methods for expressing proteins such as viral proteins, proteins required for capsid assembly, and proteins that assemble into multiprotein complexes in a manner analogous to viral capsids using a cell-free system comprising not more than about 5% wheat germ extract. Further provided are methods to assay for compounds that modulate viral protein, viral capsid assembly, and assembly of proteins into multiprotein complexes whose disruption can ameliorate bacterial, parasitic, metabolic, oncologic, immunologic, or CNS disease in a cell-free system comprising not more than about 5% wheat germ extract.

The present invention provides nucleic acids sequences, viral particles, viruses, vectors systems, host cells, kits, apparatus, and methods of evolution of a gene product of a gene of interest. The nucleic acid sequences of the invention have been developed primarily for use in evolution of biomolecules of interest. Using a split, non-competent viral vector, the gene of interest can be stably and recombinantly integrated into the viral vector via in-frame insertion of the open reading frame of the gene of interest with an aspect of the native but attenuated viral genome. This configuration of the viral genome, leveraging a split viral vector and an aspect of a viral factor that has been shown to robustly interact with the split viral vector, enables the serial passaging of the recombinant viral particle in an indel and recombination-averse manner.

Embodiments herein relate to compositions of and methods for live viruses. In certain embodiments, a live, attenuated virus composition includes, but is not limited to, one or more live, attenuated viruses and compositions to reduce inactivation and/or degradation of the live, attenuated virus. In other embodiments, the live, attenuated virus composition may be a vaccine composition. In yet other compositions, a live, attenuated virus composition may include at least one carbohydrate, at least one protein and at least one high molecular weight surfactants for reducing inactivation and/or degradation of the live, attenuated virus.

The present invention relates to the use of increased culture pH, relative to standard insect cell culture conditions, during baculovirus infection of lepidopteran insect cells to enable production of recombinant chikungunya (CHIKV) virus like particles (VLPs). The invention further relates to adapted insect cell lines derived from insect cells such as Sf21, which can grow robustly at elevated culture pH, the use of said cell lines to recombinantly produce pH sensitive proteins in the correct conformation and increase expression of recombinant proteins relative to standard insect cell lines. In some embodiments of the invention, the cells are useful for recombinant production of CHIKV VLPs. The invention also relates to a method for the production of a pH-adapted lepidopteran insect cell line. In some embodiments of said method, the cell line is produced and/or maintained in reduced phosphate serum-free insect cell media.

The invention concerns recombinant DNA's comprising cDNA of genomic RNA of a Salmonidae alphavirus preceded by a spacer sequence, under the control of a suitable promoter. Said recombinant DNAs are useful for obtaining expression vectors, producing recombinant Salmonidae alphavirus, and for obtaining vaccines.

The invention features modified alphavirus or flavivirus virus-like particles (VLPs). The invention provides methods, compositions, and kits featuring the modified VLPs. The invention also features methods for enhancing production of modified VLPs for use in the prevention or treatment of alphavirus and flavivirus-mediated diseases. The invention also provides methods for delivering agents to a cell using the modified VLPs.

The present invention relates to the discovery of a high titer hybrid-virus vector that gives rise to high titer virus like vesicles (VLVs) that can be used as a vaccine. The invention includes compositions and methods of generating an evolved hybrid-virus vector vaccine and selecting high titer VLVs, methods of treating and/or preventing or immunizing against, a specific disease or disorder, and methods of inducing a memory T cell and B cell immune response in a subject administered the VLV composition produced thereby. Furthermore, the invention encompasses a pharmaceutical composition for vaccinating a subject as well as a high titer protein expression system.