ASIALOGLICOPROTEINAS DEL VIRUS DE LA HEPATITIS C
DOS PROTEINAS DE LA CUBIERTA DEL VIRUS DE LA HEPATITIS C SE EXPRESAN SIN SIALILACION (E1 Y E2). LA EXPRESION RECOMBINANTE DE ESTAS PROTEINAS EN LOS EUCARIOTES INFERIORES, O EN LAS CELULAS DE MAMIFEROS EN LOS CUALES LA GLICOSILACION TERMINAL ESTA BLOQUEADA, DA LUGAR A PROTEINAS RECOMBINANTES QUE SON...
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Zusammenfassung: | DOS PROTEINAS DE LA CUBIERTA DEL VIRUS DE LA HEPATITIS C SE EXPRESAN SIN SIALILACION (E1 Y E2). LA EXPRESION RECOMBINANTE DE ESTAS PROTEINAS EN LOS EUCARIOTES INFERIORES, O EN LAS CELULAS DE MAMIFEROS EN LOS CUALES LA GLICOSILACION TERMINAL ESTA BLOQUEADA, DA LUGAR A PROTEINAS RECOMBINANTES QUE SON MAS PARECIDAS A LAS GLICOPROTEINAS NATIVAS DEL VHC. CUANDO SE AISLAN POR AFINIDAD A LECTINA GNA, LAS PROTEINAS E1 Y E2 SE AGREGAN COMO PARTICULAS SEMEJANTES A VIRUS. LAS CELULAS PORTADORAS DE UN RECEPTOR PARA MANOSA O UN RECEPTOR PARA ASIALOGLICOPROTEINA SON CAPACES DE SER INFECTADAS POR EL VHC Y PROMOVER EL CULTIVO DEL VIRUS.
The following are claimed: (A) an isolated hepatitis C virus (HCV) a sialoglycoprotein (AG) selected from E1 and E2; (B) a method for purifying HCV AGs, which comprises (a) contacting a compsn. contg. HCV AGs with a mannose-binding protein, e.g. concanavalin A (ConA) or Galanthus nivalus agglutinin (GNA) and (b) isolating the portion of the compsn. which binds to the mannose-binding protein, e.g. by elution with mannose; (c) an assay kit for detecting the presence of HCV AGs, comprising (a) a solid support, (b) a mannose-binding protein, and (c) an antibody specific for the HCV AG, where one of the antibody and the mannose-binding protein is bound to the solid support; (D) a method for determining exposure to or infection by HCV, in which any HCV in a sample of body fluid is concd. by contact with a mannose-binding protein prior to assay; (E) a cell transformed with a vector for recombinant expression of a HCV AG, where the vector comprises a structural gene encoding a glycosoylation signal, a HCV AG, a regulatory sequence operable in the host cell and capable of regulating expressio fo the HCV AG, and a selectable marker, where the cell does not sialylate glycoproteins; and (F) a method for reducing or eliminatng the presence of HCV in plasma, serum or other biological liquids. |
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