The present disclosure provides an automated method of producing viral vectors, utilizing engineered viral vector-producing cell lines, or packaging cells, within a fully-enclosed cell engineering system. Exemplary viral vectors that can be produced include lentivirus vectors, adeno-associated virus vectors, baculovirus vectors and retrovirus vectors.

The current teachings are directed to novel virus free cells lines derived from virus-contaminated staring material, such as an organism or a cell line. Methods for obtaining virus free cell lines obtained from virus-contaminated starting material are also provided. Exemplary virus free cell lines include: novel cell lines derived from a Spodoptera frugiperda cell line contaminated with Sf-rhabdovirus, wherein the novel cell lines lack Sf-rhabdovirus; and novel cell lines derived from a Trichoplusia ni cell line contaminated with an alphanodavirus, wherein the novel cell line lacks an alphanodavirus.

The current teachings are directed to virus free cells lines derived from virus-contaminated starting material, such as an organism or a cell line. Methods for obtaining virus free cell lines obtained from virus-contaminated starting material are also provided. Exemplary virus free cell lines include: cell lines derived from a Spodoptera frugiperda cell line contaminated with Sf-rhabdovirus, wherein the cell lines lack Sf-rhabdovirus; and cell lines derived from a Trichoplusia ni cell line contaminated with an alphanodavirus, wherein the cell line lacks an alphanodavirus.

The invention relates to a method for producing a recombinant baculovirus comprising n exogenous genes in an insect cell, by means of homologous recombination of a replication-deficient baculovirus genome and n transfer vectors, each comprising one of the n exogenous genes, n being an integer at least equal to 2.

Methods of using azide-modified biomolecules, such as fatty acids, carbohydrates and lipids, to treat a plant, an insect or an animal infected with a virus or to inhibit infectivity of a virus, such as the human immunodeficiency virus, are provided. Also provided are methods of labeling a virus, such as human immunodeficiency virus, with an azide-modified biomolecule, such as a fatty acid, a carbohydrate, or an isoprenoid lipid. Also, provided are methods of tracking a virus in vivo, with an azide-modified biomolecule, such as a fatty acid, a carbohydrate, or an isoprenoid lipid. The azide-modified biomolecules may be combined with a pharmaceutically acceptable excipient to produce a pharmaceutical composition, optionally containing another anti-viral agent and/or a delivery agent, such as a liposome.

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).

The present disclosure is related generally to systems and methods for high level expression of recombinant proteins from baculovirus in insect cells. In particular, the methods and systems described herein allow for high levels of baculovirus production in insect cells and/or high levels of protein production in insect cells using a chemically-defined, yeast lysate-free insect cell medium. The disclosure also relates to compositions and kits for culturing, transfecting, and/or producing recombinant protein in insect cells.

Method for preparing, in an insect cell, a recombinant baculovirus comprising one or more transgene(s) each encoding a protein maturation enzyme and n transgenes each encoding a polypeptide of interest, by homologous recombination between a replication deficient baculovirus genome which comprises one or more transgene(s) each encoding a protein maturation enzyme and n transfer vectors each comprising one of the n transgenes each encoding a polypeptide of interest, n being an integer at least equal to 2.

Two new Helicoverpa armigera single nucleopolyhedrovirus genotypes, HearSNPV, HearSNPV-SP1B and HearSNPV-LB6, each originating from mixtures of genotypes obtained from different locations and crops, are described. Each exhibits specific insecticidal activity against H. armigera larvae comparable to that of commonly used commercial insecticides. Further, mixing the two genotypes, especially in the ratio of 1:1, within co-occluded virions of the mixed genotypes, is capable of controlling H. armigera infestations of tomato crops and is as efficacious as commonly used chemical and biological insecticides. Their use as bioinsecticides is safe for vertebrates, in that they specifically target arthropods. In addition, they are easy to produce and good yields can be obtained by orally inoculating H. armigera larvae with HearSNPV occlusion bodies.

Production of virus occlusion bodies that occlude virions comprising genomes of different species of baculoviruses that can be used to combat insect pests. A method is presented for the production of occlusion derived virions (ODVs) that simultaneously comprise genomes of different baculovirus species, occluded in a viral occlusion body (OB) with the structural and morphological features characteristic of baculoviruses. Mixed genome ODVs and OBs can be produced by co-infecting insect cells or insect hosts using two or more different baculoviruses species. Co-infection may be achieved by simultaneous inoculation of the different baculoviruses or with a time interval between inoculations, which results in different proportions of each species' genomes in the ODVs and OBs that are produced. The produced OBs can be used either directly for preparing an insecticide, or to infect susceptible insects to produce larger quantities of mixed genome ODVs and OBs, also useful for combating pest insects.