Methods of purifying virus-like particles (VLPs) that are substantially free of process contaminants and infectious agents. The methods incorporate, for example, low-pH treatment during harvest and/or inactivation by a solvent and/or detergent during VLP capture.

The present disclosure provides viral microparticles comprising genetically-engineered baculoviruses (at least partially) embedded in a polymeric matrix for the local delivery of therapeutic nucleic acid molecules to the cells of a vertebrate individual (optionally in combination with a medical implant such as vascular stent platform). The viral microparticles are especially useful for promoting the healing of a wound as well as the repair of a blood vessel and prevent pathological scarring. Also provided herein are processes for making the viral microparticles, pharmaceutical compositions comprising viral microparticles as well as supports comprising the viral microparticles for the locating the viral microparticles in a wound or in the vicinity of a wound.

The present disclosure provides vaccine compositions for prophylaxis and treatment of Zika virus infections comprising Zika virus antigens in immunogenic compositions, and in combination of Zika antigens with one or more arbovirus antigens such as Chikungunya virus and Japanese encephalitis virus antigens, methods of preparation and production of such compositions for use as vaccines for eliciting immune response in mammals against the above mentioned pathogens.

Compositions and methods are disclosed for producing adeno-associated virus (AAV) in insect cells in vitro. Recombinant baculovirus vectors include an AAV Capsid gene expression cassette (Cap), an AAV Rep gene expression cassette (Rep), and a baculovirus homologous region (hr) located up to about 4 kb from a start codon in an AAV expression cassette. Production levels of baculovirus and AAV in insect cells harboring recombinant baculovirus comprising a Cap, a Rep, and an hr are higher compared to controls comprising a Cap and a Rep but no hr. Furthermore, levels of baculovirus and AAV production in insect cells infected with recombinant baculovirus comprising a Cap, a Rep, and an hr are comparatively stable over serial passages of cells, whereas levels of baculovirus and AAV production decline over serial passages of insect cells comprising recombinant baculovirus comprising a Cap and a Rep, but no hr.

The present disclosure presents methods for producing baculovirus infected insect cells (BIICs). The present disclosure describes methods and systems for use in the production of adeno-associated virus (AAV) particles, compositions and formulations, including recombinant adeno-associated viruses (rAAV). In certain embodiments, the production process and system use Baculoviral Expression Vectors (BEVs) and/or Baculoviral Infected Insect Cells (BIICs) in the production of rAAVs. In certain embodiments, the present disclosure presents methods and systems for designing, producing, clarifying, purifying, formulating, filtering and processing rAAVs and rAAV formulations. In certain embodiments, the production process and system use Spodoptera frugiperda insect cells (such as Sf9 or Sf21) as viral production cells (VPCs).

Methods of determining the stoichiometry of a viral capsid and/or determining the heterogeneity of protein components in a viral capsid are disclosed.

The present disclosure relates to a recombinant baculovirus expression vector (rBEV) for the production of closed-ended DNA (ceDNA) in insect cells.

The design, assembly, and use of novel sequences comprising targeting and insertion sites for site-specific bacterial transposons are disclosed. One aspect relates to a nucleotide sequence comprising an attachment site for a site-specific transposon operably-linked to a screenable or selectable marker sequence, wherein said marker sequence encodes one or more active or inactive polypeptides capable of conferring a screenable or selectable phenotype upon a cell comprising the marker sequence, wherein insertion of the site-specific transposon into the attachment site changes the phenotype of a cell comprising the screenable or selectable marker sequence. High and low copy number vectors comprising the sequences, designated synthemids, including plasmids capable of propagating in bacteria, and shuttle vectors, capable of propagating in bacteria and a eukaryotic host cell or two types of bacteria by means of distinct replicons, are also disclosed. Related aspects include the design and assembly of synthetic insect and mammalian virus shuttle vectors, including shuttle vectors comprising segments of a double-stranded DNA virus, such as a baculovirus, which propagates in insect cells, or a herpesvirus, an adenovirus, or a pox virus, which propagate in mammalian cells. Other aspects relate to use of modified vectors to express polypeptides for use as therapeutic drug products, as vaccines, or as components of cell or gene therapy vector systems, and in model and crop plant cells, tissues, and whole plants to facilitate the basic and applied studies leading to improved food products, and as tools advancing the interests of institutions involved in industrial and environmental biotechnology.

The present invention provides a respiratory syncytial virus (RSV) recombinant fusion protein (F protein) in which a polymerization domain derived from a foreign protein is bound to the C terminal of a fusion protein (F protein) lacking a transmembrane domain of a wild-type respiratory syncytial virus (RSV) fusion protein (F protein). The recombinant fusion protein of the present invention is soluble and can retain an F protein trimer. Excellent immune-inducing effects can be expected from the recombinant fusion protein of the present invention, and vaccine composition containing same.

The present invention is directed to the expression and secretion the Zika virus envelope protein. Elements of the pre-membrane and envelope sequence have been modified to enhance the expression of the envelope protein as a secreted product in the culture medium of transformed insect cell lines. The expressed and purified product is suitable as a vaccine antigen.