Expression of heterologous proteins
AssigneeChiron Corporation
InventorCesira Galeotti
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Abstract
An expression system which provides heterologous proteins expressed by a non-native host organism but which have native-protein-like biological activity and/or structure. Disclosed are vectors, expression hosts and methods for expressing the heterologous proteins. The expression system involves co-expression of protein factor(s) which is/are capable of catalyzing disulphide bond formation and desired heterologous protein(s). The expression system is presented using yeast cells as the preferred host, protein disulphide isomerase (PDI) and thioredoxin (TRX) as the preferred examples of the protein factors and HCV-E2715 envelope glycoprotein and human FIGF as the preferred examples of the heterologous proteins.
Claims28
A vector comprising:
- (i) an expression cassette comprising a DNA sequence encoding thioredoxin (TRX);
- and
- (ii) an expression cassette comprising DNA sequence(s) encoding one or more heterologous proteins;
- wherein said one or more heterologous proteins do(es) not form fusion protein(s) with said TRX.
A vector comprising:
- (i) an expression cassette comprising a DNA sequence encoding a protein that catalyzes disulphide bond formation;
- and
- (ii) an expression cassette comprising DNA sequence(s) encoding a heterologous protein selected from the group consisting of hepatitis C virus (HCV) E2715 envelope glycoprotein and human c-fos-induced growth factor (FIGF);
- wherein said heterologous protein, when expressed, is localised in the endoplasmic reticulum (ER) and/or other secretory compartments.
A host organism transformed with:
- (i) one or more vectors comprising an expression cassette comprising a DNA sequence encoding a protein that catalyzes disulphide bond formation;
- and
- (ii) one or more vectors comprising an expression cassette comprising DNA sequence(s) encoding a heterologous protein selected from the group consisting of hepatitis C virus (HCV) E2715 envelope glycoprotein and human c-fos-induced growth factor (FIGF);
- wherein said heterologous protein, when expressed, is localized in the ER and/or other secretory compartments.
A method of producing a transformed organism, said method comprising:
- (i) transforming a host organism with one or more vectors comprising an expression cassette comprising a DNA sequence encoding a protein that catalyzes disulphide bond formation;
- and
- (ii) either simultaneously or sequentially further transforming said host organism wich one or more vectors comprising an expression cassette comprising a DNA sequence encoding a heterologous protein selected from the group consisting of hepatitis C virus (HCV) E2715 envelope glycoprotein and human c-fos-induced growth factor (FIGF);
- wherein said heterologous protein, when expressed, is localized in the ER and/or other secretory compartments.
Dependencies are inferred from the claim text.