Chemistry: molecular biology and microbiology – Vector – per se
Reexamination Certificate
2005-09-13
2005-09-13
Guzo, David (Department: 1636)
Chemistry: molecular biology and microbiology
Vector, per se
C435S236000, C435S239000, C435S455000, C435S456000, C435S457000, C424S093200, C424S199100
Reexamination Certificate
active
06943015
ABSTRACT:
Large-scale packaging of alphavirus replicons is accomplished by co-transfecting host cells with three RNA molecules: (1) an alphavirus replicon comprising a sequence encoding a heterologous protein; (2) a helper RNA encoding a capsid protein and (3) a helper RNA encoding two alphavirus glycoproteins. The helper RNAs contain cis-acting elements that allow efficient replication of the helper RNAs and their packaging into viral particles as well as packaging of replicon genomes. These populations of viral particles can be propagated at high titers in cell culture by infecting cells at high multiplicity. Propagation of packaged replicons at an escalating scale is useful for large-scale production of recombinant proteins and/or vaccine.
REFERENCES:
Frolov, I. et al., “Selection of RNA Replicons Capable of Persistent Noncytopathic Replication of Mammalian Celss”, 1999, J. Virol. vol. 73: pp. 3854-3865.
Polo, J.M. et al., “Stable alphavirus packaging cell lines for Sindbis virus- and Semliki Forest virus-derived vectors”, 1999, PNAS, vol. 96: pp. 4598-4603.
Smerdou, C. et al., “Two-Helper RNA System for Production of Recombinant Semliki Forest Virus Particles”, J. Virol., vol. 73: pp. 1092-1098.
Frolov Ilya
Frolova Elena
Adler Benjamin Aaron
Burkhart Michael
Guzo David
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