Recombinant hepatitis c virus rna replicase

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/54 (2006.01) C12N 9/12 (2006.01) C12Q 1/48 (2006.01) C12Q 1/68 (2006.01) C12Q 1/70 (2006.01) G01N 33/569 (2006.01) G01N 33/573 (2006.01)

Patent

CA 2233309

A recombinant RNA-dependent RNA polymerase of hepatitis C virus (r-HCV-RDRP) coding DNA was cloned and expressed yielding active enzyme in vitro. The r-HCV- RDRP can include up to 20 added amino acids and up to nine deleted or substituted amino acids at the NH2-terminus of the encoded amino acid sequence. The invention provides method to solubilize r-HCV-RDRP from a host cell lysate and purified r-HCV-RDRP. Methods for screening for inhibitors of r- HCV-RDRP in vitro, for making stably transfected mammalian cells expressing r- HCV-RDRP and for in vivo testing of r-HCV-RDRP inhibitors in vivo are disclosed. The invention provides antibodies to r-HCV-RDRP and methods for detecting antibodies to HCV-RDRP in serum of human patients.

L'invention concerne une ARN polymérase ARN-dépendante recombinée de l'ADN codant le virus de l'hépatite C (r-HCV-RDRP) qui, une fois clonée et exprimée, produit in vitro une enzyme active. Le r-HCV-RDPR peut comprendre jusqu'à 20 acides aminés ajoutés, et jusqu'à 9 acides aminés supprimés ou substitués au niveau de la terminaison NH¿2? de la séquence acide aminée codée. L'invention concerne également un procédé pour solubiliser r-HCV-RDRP à partir d'un lysat de cellules hôtes, et r-HCV-RDRP purifié. L'invention porte en outre sur des procédés de criblage d'inhibiteurs de r-HCV-RDRP in vitro; de fabrication de cellules mammaliennes transfectées de manière stable, exprimant r-HCV-RDRP; et de détection $i (in vivo) d'inhibiteurs de r-HCV-RDRP in vivo. L'invention porte enfin sur des anticorps contre r-HCV-RDRP et des procédés de détection d'anticorps contre HCV-RDRP dans le sérum de patients humains.

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