Methods for the preparation of hepatitis C virus multiple copy epitope fusion antigens

US6514731B1 · kind B1 · utility

Assignee

• Chiron Corporation · US

Inventors

• Pablo D. T. Valenzuela · San Francisco, US
• David Chien · Emeryville, US

Key dates

• Filing date: May 24, 1996
• Grant date: Feb 4, 2003
• Expiry date: May 24, 2016

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2770/24222
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Human hepatitis C virus (HCV) has been identified as the aetiological agent of non-A, non-B hepatitis (NANBH). HCV viruses display considerable genotypic and phenotypic heterogeneity. Thus, there is considerable need in the art for more sensitive reagents that facilitate the detection of HCV variants. The genome of hepatitis C virus (HCV) consists of seven functional regions: the core, E1, E2/NS1, NS2, NS3, NS4, and NS5 regions. An attempt was made to improve the sensitivity of anti-HCV assays by developing multiple copy epitope fusion antigens (MEFAs) which incorporate the major immunodominant epitopes from the functional regions of the HCV genome. These MEFAs are encompassed by the following generic structural formula: (A)x—(B)y—(C)z. This formula represents a linear amino acid sequence comprising multiple copies of one HCV epitope (A) linked to multiple copies of another HCV epitope (B) which in turn is linked to multiple copies of yet another HCV epitope (C). Expression vectors carrying nucleic acid sequences comprising MEFA antigens carrying multiple copies of epitopes derived from the viral core, E1, E2, NS3, NS4, and NS5 regions were prepared. The resultant MEFA …