Innogenetics N.v.

N.v. Innogenetics S.a.

Fons Bosman

Geert Maertens

Erik Depla

Guy De Martynoff

Marie-Ange Buyse

Erwin Sablon

Geert Deschamps

Isabelle Samson

Annie Van Broekhoven

Manfred Suckow

Gert Verheyden

Joost Haelewyn

Marie Ange Buyse

The current invention relates to vectors and methods for efficient expression of HCV envelope proteins in eukaryotic cells. More particularly said vectors comprise the coding sequence for an avian lysozyme signal peptide or a functional equivalent thereof joined to a HCV envelope protein or a part thereof. Said avian lysozyme signal peptide is efficiently removed when the protein comprising said avian lysozyme signal peptide joined to a HCV envelope protein or a part thereof is expressed in a eukaryotic cell. Suitable eukaryotic cells include yeast cells such asorcells.

Constructs and methods for expression of recombinant HCV envelope proteins

The current invention relates to HCV envelope proteins or parts thereof which are the product of expression in eukaryotic cells. More particularly said HCV envelope proteins are characterized in that on average up to 80% of their N-glycosylation sites are core-glycosylated. Of these N-glycosylated sites more than 70% are glycosylated with an oligomannose having a structure defined by Man(8 to 10)-GlcNAc(2). Furthermore, the ratio of the oligomannose with structure Man(7)-GlcNAc(2) over the oligomannose with structure Man(8)-GlcNAc(2) is less than or equal to 0.45. Less than 10% of the oligomannoses is terminated with an &#x3b1;1,3 linked mannose. The HCV envelope proteins of the invention are particularly suited for diagnostic, prophylactic and therapeutic purposes. A suitable eukaryotic cell for production of the HCV envelope proteins of the invention is acell.

Core-glycosylated HCV envelope proteins

The present invention relates to a method for purifying recombinant HCV single or specific oligomeric envelope proteins selected from the group consisting of E1 and/or E2 and/or E1/E2, characterized in that upon lysing the transformed host cells to isolate the recombinantly expressed protein a disulphide bond cleavage or reduction step is carried out with a disulphide bond cleavage agent. The present invention also relates to a composition isolated by such a method. The present invention also relates to the diagnostic and therapeutic application of these compositions. Furthermore, the invention relates to the use of HCV E1 protein and peptides for prognosing and monitoring the clinical effectiveness and/or clinical outcome of HCV treatment

Purified hepatitis C virus envelope proteins for diagnostic and therapeutic use

The present invention relates to a method for purifying recombinant HCV single or specific oligomeric envelope proteins selected from the group consisting of E1 and/or E2 and/or E1/E2, characterized in that upon lysing the transformed host cells to isolate the recombinantly expressed protein a disulphide bond cleavage or reduction step is carried out with a disulphide bond cleavage agent. The present invention also relates to a composition isolated by such a method. The present invention also relates to the diagnostic and therapeutic application of these compositions. Furthermore, the invention relates to the use of HCV E1 protein and peptides for prognosing and monitoring the clinical effectiveness and/or clinical outcome of HCV treatment.

The present invention relates to the general field of recombinant protein expression, purification of recombinant proteins, diagnosis of HCV infection, prophylactic treatment against HCV infection and to the prognosing/monitoring of the clinical efficiency of treatment of an individual with chronic hepatitis, or the prognosing/monitoring of the natural disease. In particular, the present invention relates to the use of yeast, i.e.orglycosylation minus strains, for the efficient expression of HCV envelope proteins that are core-glycosylated, purification methods for these proteins, and the use in various applications, such as the use in diagnosis, prophylaxis or therapy of HCV envelope proteins purified according to the present invention.

Expression of core-glycosylated HCV envelope proteins in yeast

The present invention relates to a method for purifying recombinant HCV single or specific oligomeric envelope proteins selected from the group consisting of E1 and/or E1/E2 characterized in that upon lysing the transformed host cells to isolate the recombinantly expressed protein a disulphide bond cleavage or reduction step is carried out with a disulphide bond cleavage agent. The present invention also relates to a composition isolated by such a method. The present invention also relates to the diagnostic and therapeutic application of these compositions. Furthermore, the invention relates to the use of HCV E1 protein and peptides for prognosing and monitoring the clinical effectiveness and/or clinical outcome of HCV treatment.

The present invention relates to a method for purifying recombinant HCV single or specific oligomeric envelope proteins selected from the group consisting of E1 and/or E1/E2, characterized in that upon lysing the transformed host cells to isolate the recombinantly expressed protein a disulphide bond cleavage or reduction step is carried out with a disulphide bond cleavage agent. The present invention also relates to a composition isolated by such a method. The present invention also relates to the diagnostic and therapeutic application of these compositions. Furthermore, the invention relates to the use of HCV E1 protein and peptides for prognosing and monitoring the clinical effectiveness and/or clinical outcome of HCV treatment.

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Opwijk, Belgium

Fons Bosman