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FEBS Lett. 2013 Oct 11;587(20):3335-40. doi: 10.1016/j.febslet.2013.07.057. Epub 2013 Sep 7.

Structural and functional characterization of the single-chain Fv fragment from a unique HCV E1E2-specific monoclonal antibody.

Author information

1
Univ. Grenoble Alpes, Unit for Virus Host-Cell Interactions, F-38000 Grenoble, France; CNRS, Unit for Virus Host-Cell Interactions, F-38000 Grenoble, France; Unit for Virus Host-Cell Interactions, Univ. Grenoble Alpes-EMBL-CNRS, 6 rue Jules Horowitz, 38042 Grenoble, France.

Abstract

The nucleotide sequence of the unique neutralizing monoclonal antibody D32.10 raised against a conserved conformational epitope shared between E1 and E2 on the serum-derived hepatitis C virus (HCV) envelope was determined. Subsequently, the recombinant single-chain Fv fragment (scFv) was cloned and expressed in Escherichia coli, and its molecular characterization was assessed using multi-angle laser light scattering. The scFv mimicked the antibody in binding to the native serum-derived HCV particles from patients, as well as to envelope E1E2 complexes and E1, E2 glycoproteins carrying the viral epitope. The scFv D32.10 competed with the parental IgG for binding to antigen, and therefore could be a promising candidate for therapeutics and diagnostics.

KEYWORDS:

3-(1-pyridino)-1-propanesulfonates; Anti-HCV therapy; Antibody engineering; CDR; DAAs; E1E2; ELISA; FR; GT; HCV; HCVsp; HRP; Hepatitis C virus; IMAC; IPTG; IgG; LB medium; Leibovitz medium; MALLS; NDSB; NR; PCR; PEG-IFN; PVDF; R; RBV; SDS–PAGE; SOC; Single chain Fv fragment; TBS; Tris buffer saline; V(H); V(L); complementarity-determining region; direct-acting antivirals; enzyme-linked immunoabsorbent assay; framework region; genotype; heavy chain variable region; hepatitis C virus; horseradish peroxidase; immobilized metal affinity chromatography; immunoglobulin G; isopropylthio-β-galactoside; light chain variable region; mAb; monoclonal antibody; multi-angle laser light scattering; non-reducing; pegylated interferon-α; polymerase chain reaction; polyvinylidene difluoride; reducing; ribavirin; scFv; serum-derived HCV particles; single chain antibody fragment; sodium dodecylsulfate–polyacrylamide gel electrophoresis; standard of care

PMID:
24021643
DOI:
10.1016/j.febslet.2013.07.057
[Indexed for MEDLINE]
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