Disclosed are methods of producing poxvirus-based vectors or recombinant poxvirus-based vectors from naturally derived, chemically synthesized DNA fragments, or a combination of naturally derived and chemically synthesized DNA fragments. One or more DNA sequences encoding one or more antigens, subunits or fragments thereof or other heterologous gene sequences are inserted in one or more poxvirus insertion sites in one or more DNA fragments. The methods include transfecting a host cell with one or more circular or linear DNA fragments such that a poxvirus or recombinant poxvirus is reconstituted in the host cell, the reconstituted poxvirus or recombinant poxvirus comprising the genome of a desired poxvirus. Also disclosed are poxviruses or recombinant poxviruses produced by the technology and uses thereof.

The present invention generally relates to a process for designing a recombinant poxvirus for a therapeutic vaccine, i.e. personalized cancer vaccine, said recombinant poxvirus comprising one or more expression cassettes, each for expression of a fusion of a plurality of peptides, i.e. neopeptides, characterized in that it comprises performing by processing means (11) of a server (1) the steps of : (a) selecting a first subset of candidate peptides, wherein said peptides present transmembrane scores below a TMS threshold; b) determining an optimal distribution of the candidate peptides from said first subset to the expression cassette(s) among a plurality of possible distributions, wherein said optimal distribution presents, if there are at least two expression cassettes, the lowest range between the hydropathy scores of at least two expression cassettes; (c) for each expression cassette, determining an optimal slot allocation of the candidate peptides as function of cassette slot occupancy rule so as to select the peptide fusion with the lowest TM score; (d) determining a DNA transfer sequence comprising the nucleotide sequence of the one or more expression cassette(s) for generation of said recombinant poxvirus.

The present invention relates to the development and manufacturing of viral vaccines. In particular, the invention relates to the field of industrial production of viral vectors and vaccines, more in particular to the use of avian embryonic stem cells, preferably the EBx® cell line derived from duck embryonic stem cells, for the production of viral vectors and viruses. The invention is particularly useful for the industrial production of viral vaccines to prevent viral infection of humans and animals.

Expression vectors ideal for use in vaccinating individuals against disease based on vaccinia virus and other chordopoxviruses having high expression of recombinant genes and low expression of vector genes in target animals, and low expression of recombinant genes and high expression of vector genes in cells used for propagation.

The present invention relates to a method for purifying an enveloped virus. The present invention further relates to an enveloped virus or a plurality of enveloped viruses obtainable by said method.

The present invention provides a method for producing a virus and a harvesting solution composition. The method includes culturing cells, wherein the cells have been inoculated with viruses or have been transfected with viral packaging elements; and contacting the cultured cells with a harvesting solution composition to harvest the viruses by one-step, wherein the harvesting solution composition comprises a trypsin, a pH buffer and optionally a nuclease, and the pH of the harvesting solution composition is greater than 7.5 and no more than 10.5. The virus production method of the present invention has advantages of simple operation, easy scale-up, stable yield and so on, and the yield is unexpectedly and significantly improved compared to the prior art, and it can ensure the integrity of the viral particles without damaging the biological activity of the viruses. Therefore, it is very suitable for large-scale production of viruses.

Certain embodiments are directed to viral production processes for the large scale production of vaccinia virus.

The present invention relates to oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine and to methods for cultivating cells in said oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine. The invention also relates to methods for expressing at least one protein in a medium comprising at least 0.5 mg/L of a polyamine and to methods for producing at least one virus in a medium comprising at least 0.5 mg/L of a polyamine.

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.

Provided are compounds of Formula (II) that enhance the efficacy of viruses by increasing spread of the virus in cells, increasing the titer of virus in cells, or increasing the antigen expression from a virus, gene or trans-gene expression from a virus, or virus protein expression in cells. Other uses, compositions and methods of using same are also provided. ##STR00001##