title: Expression of a functional scFv fragment of an anti-idiotypic antibody with a beta-lactam hydrolytic activity.
abstract: The single chain variable fragment (scFv) of an anti-idiotypic catalytic monoclonal antibody, 9G4H9, displaying a beta-lactamase-like activity was cloned. The recombinant protein was expressed through the periplasm in Escherichia coli in the presence or in the absence of FkpA, a chaperone-like enzyme and tested for its hydrolytic activity. The results show that the catalytic parameters for hydrolysis of ampicillin by scFv9G4H9 are clearly influenced by the presence of FkpA, indicating that the correct folding of the fragment represents a crucial step for catalysis.
authors.affiliation: Génie Enzymatique et Cellulaire, UMR 6022 CNRS, Université de Technologie de Compiègne, BP 20529, 60205 Compiègne Cedex, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Padiolleau-Lefèvre
authors.forename: Hélène
authors.initials: H
authors.lastname: Débat
authors.forename: Denis
authors.initials: D
authors.lastname: Phichith
authors.forename: Daniel
authors.initials: D
authors.lastname: Thomas
authors.forename: Alain
authors.initials: A
authors.lastname: Friboulet
authors.forename: Bérangère
authors.initials: B
authors.lastname: Avalle
mesh_terms: D000888:Antibodies, Anti-Idiotypic / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017151:Antibodies, Catalytic / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001665:Binding Sites; D003001:Cloning, Molecular; D006868:Hydrolysis; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008958:Models, Molecular; D018716:Molecular Mimicry; D001618:beta-Lactamases / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D000888:Antibodies, Anti-Idiotypic; D017151:Antibodies, Catalytic; D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments; D001618:beta-Lactamases