Expression of a functional scFv fragment of an anti-idiotypic antibody with a beta-lactam hydrolytic activity.
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.
Padiolleau-Lefèvre
Séverine
S
Génie Enzymatique et Cellulaire, UMR 6022 CNRS, Université de Technologie de Compiègne, BP 20529, 60205 Compiègne Cedex, France.
Débat
Hélène
H
Phichith
Denis
D
Thomas
Daniel
D
Friboulet
Alain
A
Avalle
Bérangère
B
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*
D000888:Antibodies, Anti-Idiotypic; D017151:Antibodies, Catalytic; D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments; D001618:beta-Lactamases