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Structural and functional characterization of the single‐chain Fv fragment from a unique HCV E1E2‐specific monoclonal antibody
Author(s) -
Fallecker Catherine,
Tarbouriech Nicolas,
Habib Mohammed,
Petit Marie-Anne,
Drouet Emmanuel
Publication year - 2013
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2013.07.057
Subject(s) - epitope , monoclonal antibody , recombinant dna , microbiology and biotechnology , conformational epitope , virology , epitope mapping , antibody , glycoprotein , immunoglobulin light chain , chemistry , phage display , viral envelope , linear epitope , biology , biochemistry , gene , genetics
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.