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US 12,365,879 B2Granted
Synthetic chimeric poxviruses
AssigneeTonix Pharma Limited
InventorsDavid Evans, Ryan Noyce, Seth Lederman
Life of this patent
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Abstract
The invention relates, in general, to synthetic chimeric poxviruses, compositions comprising such viruses, and the development and use of systems and methods for producing such synthetic chimeric poxviruses. The synthetic chimeric poxviruses are well suited for live virus vaccines and pharmaceutical formulations.
Claims20
A method of producing a synthetic chimeric orthopoxvirus (scOPV) comprising the steps of:
- (i) chemically synthesizing overlapping DNA fragments that correspond to substantially all of a wild type genome selected from the group consisting of a camelpox virus, a cowpox virus, an ectromelia virus, a horsepox virus, a monkeypox virus, a vaccinia virus, a variola virus, a rabbitpox virus, a raccoon poxvirus, a skunkpox virus, a Taterapox virus, an Uasin Gishu disease virus, and a volepox virus;
- (ii) chemically synthesizing terminal hairpin loops independently derived from an orthopoxvirus and ligating the terminal hairpin loops derived from the chemically synthesized hairpin loops to the left and right termini of the overlapping DNA fragments derived from the chemically synthesized DNA of step (i), and wherein the orthopoxvirus is selected from the group consisting of a camelpox virus, a cowpox virus, an ectromelia virus, a horsepox virus, a monkeypox virus, a vaccinia virus, a variola virus, a rabbitpox virus, a raccoon poxvirus, a skunkpox virus, a Taterapox virus, an Uasin Gishu disease virus, and a volepox virus;
- (iii) transfecting the overlapping DNA fragments derived from the overlapping chemically synthesized DNA fragments of step (i) and the ligated terminal hairpin loops of step (ii) into helper virus-infected cells;
- (iv) culturing said cells to produce a mixture of helper virus and scOPV particles in said cells;
- and
- (v) plating the mixture on OPV-specific host cells to recover the scOPV.
3 more elements
A method of producing a synthetic chimeric horsepox virus (scHPXV) comprising the steps of:
- (i) chemically synthesizing overlapping DNA fragments that correspond to substantially all of a wild type HPXV genome;
- (ii) chemically synthesizing terminal hairpin loops independently derived from an orthopoxvirus and ligating the terminal hairpin loops derived from the chemically synthesized hairpin loops to the left and right termini of the overlapping DNA fragments derived from the chemically synthesized DNA of step (i), and wherein the orthopoxvirus is selected from the group consisting of a camelpox virus, a cowpox virus, an ectromelia virus, a horsepox virus, a monkeypox virus, a vaccinia virus, a variola virus, a rabbitpox virus, a raccoon poxvirus, a skunkpox virus, a Taterapox virus, an Uasin Gishu disease virus, and a volepox virus;
- (iii) transfecting the overlapping DNA fragments derived from the overlapping chemically synthesized DNA fragments of step (i) and the ligated terminal hairpin loops of step (ii) into helper virus-infected cells;
- (iv) culturing said cells to produce a mixture of helper virus and scHPXV particles in said cells;
- and
- (v) plating the mixture on HPXV-specific host cells to recover the scHPXV.
3 more elements
Dependencies are inferred from the claim text.