Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10871135 | FEBS Letters | 2013 | 6 Pages |
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
LB mediumPEG-IFNIMACHRPTBSRBVpegylated interferon-αscFvIPTGCDRDAAsPVDFIgGMALLSmAbNDSBSingle chain Fv fragmentisopropylthio-β-galactosideSDS–PAGEMonoclonal antibodyenzyme-linked immunoabsorbent assaystandard of caresodium dodecylsulfate–polyacrylamide gel electrophoresisimmunoglobulin GTris buffer salineELISAdirect-acting antiviralspolyvinylidene difluorideRibavirinSOCsingle chain antibody fragmentComplementarity-determining regionLight chain variable regionHeavy chain variable regionframework regionAntibody engineeringHepatitis C virusHCVpolymerase chain reactionPCRHorseradish peroxidasemulti-angle laser light scatteringGenotypeReducingimmobilized metal affinity chromatography
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Authors
Catherine Fallecker, Nicolas Tarbouriech, Mohammed Habib, Marie-Anne Petit, Emmanuel Drouet,