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Jan 29 2017

Imprimer ce Article

Multi-target Selection of Catalytic Antibodies wih β-lactamase Activity using Phage Display

Authors

Melody A. Shahsavarian, Nancy Chaaya, Narciso Costa, Didier Boquet, Alexandre Atkinson, Bernard Offmann, Srini V. Kaveri, Sébastien Lacroix-Desmazes, Alain Friboulet, Bérangère Avalle, Séverine Padiolleau-Lefèvre

Journal

FEBS Journal (Accepted Manuscript. doi:10.1111/febs.14012)

Abstract

β-lactamase enzymes responsible for bacterial resistance to antibiotics are among the most important health threats to the human population today. Understanding the increasingly vast structural motifs responsible for the catalytic mechanism of β-lactamases will help improve the future design of new generation antibiotics and mechanism-based inhibitors of these enzymes. Here we report the construction of a large murine scFv phage display library of size 2.7×109 with extended diversity by combining different mouse models. We have used two molecularly different inhibitors of the R-TEM β-lactamase as targets for selection of catalytic antibodies with β-lactamase activity. This novel methodology has led to the isolation of 5 antibody fragments, which are all capable of hydrolyzing the β-lactam ring. Structural modeling of the selected scFv has revealed the presence of different motifs in each of the antibody fragments potentially responsible for their catalytic activity. Our results confirm i) the validity of using our two target inhibitors for the in vitro selection of catalytic antibodies endowed with β-lactamase activity, and ii) the plasticity of the β-lactamase active site responsible for the wide resistance of these enzymes to clinically available inhibitors and antibiotics.

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