PMID: 7515376Feb 1, 1994Paper

Isolation and epitope characterization of human monoclonal antibodies to hepatitis C virus core antigen

Hybridoma
K SiemoneitB Subanek

Abstract

In this study we describe the establishment of two hybridoma cell lines secreting human monoclonal antibodies to the 22-kD nucleocapsid protein (core, p22) of the hepatitis C virus (HCV). For this purpose we isolated B lymphocytes from an anti-HCV positive blood donor and infected them with Epstein-Barr (EBV). We obtained several lymphoblastoid cell clones secreting antibodies to the recombinant HCV core protein. The B-cell cultures were oligoclonally expanded and two of them were fused with the (mouse:human) heteromyeloma cell line K6H6/B5. The resulting stable hybridomas produce antibodies of the IgG1/kappa (U1/F10) and the IgM/kappa (Ul/F11) isotype reacting specifically with the recombinant core protein p22. To identify the epitopes recognized by these antibodies we synthesized overlapping peptides (13-mer and 6-mer) from the amino terminus of the core amino acid sequence. Antibody reactivity to these peptides was analyzed in an immunoblot assay. Finally, we were able to define a linear epitope recognized by the Ul/F10 antibody on the nucleocapsid protein. The antibody shows specificity to the sequence N-VYLLPR-C, which corresponds to the amino acids 34-39 of the core sequence.

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Citations

Mar 1, 1996·Journal of Medical Virology·D MoradpourJ R Wands
Apr 28, 2006·Molekuliarnaia biologiia·E A RechkinaA A Kushch
Dec 13, 2005·Molecular Immunology·Yuanding ChenWenlin Yu
Nov 5, 2019·Science Immunology·Megan Sykes, David H Sachs
Feb 15, 2000·The British Journal of Surgery·B SoinP J Friend
Aug 2, 2008·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·K KiernanM Kearns-Jonker
Oct 30, 2008·Xenotransplantation·Guerard W ByrneChristopher G A McGregor
Mar 2, 2011·The Journal of Immunology : Official Journal of the American Association of Immunologists·Kim M WigglesworthUri Galili
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Mar 19, 2002·Xenotransplantation·Katsuhito TeranishiDavid K C Cooper
Feb 18, 2003·Xenotransplantation·Hideki Ohdan, Megan Sykes
Feb 8, 2003·The Journal of Immunology : Official Journal of the American Association of Immunologists·Renée MénezEnrico A Stura

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