title: De novo isolation of antibodies with pH-dependent binding properties.
abstract: pH-dependent antibodies are engineered to release their target at a slightly acidic pH, a property making them suitable for clinical as well as biotechnological applications. Such antibodies were previously obtained by histidine scanning of pre-existing antibodies, a labor-intensive strategy resulting in antibodies that displayed residual binding to their target at pH 6.0. We report here the de novo isolation of pH-dependent antibodies selected by phage display from libraries enriched in histidines. Strongly pH-dependent clones with various affinity profiles against CXCL10 were isolated by this method. Our best candidate has nanomolar affinity for CXCL10 at pH 7.2, but no residual binding was detected at pH 6.0. We therefore propose that this new process is an efficient strategy to generate pH-dependent antibodies.
authors.affiliation: a NovImmune SA ; Plan-les-Ouates ; Geneva , Switzerland.
authors.forename: Pauline
authors.initials: P
authors.lastname: Bonvin
authors.forename: Sophie
authors.initials: S
authors.lastname: Venet
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Fontaine
authors.forename: Ulla
authors.initials: U
authors.lastname: Ravn
authors.forename: Franck
authors.initials: F
authors.lastname: Gueneau
authors.forename: Marie
authors.initials: M
authors.lastname: Kosco-Vilbois
authors.forename: Amanda Ei
authors.initials: AE
authors.lastname: Proudfoot
authors.forename: Nicolas
authors.initials: N
authors.lastname: Fischer
mesh_terms: D054357:Chemokine CXCL10 / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015202:Protein Engineering; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000235:genetics
chemical_list: C114753:CXCL10 protein, human; D054357:Chemokine CXCL10; D057127:Single-Chain Antibodies
keywords: BLI, bio-layer interferometry; CDR, complementary determining region; CDRH, CDR of the heavy chain; CDRL, CDR of the light chain; ELISA, enzyme-linked immunosorbent assay; GPCR, G protein-coupled receptor; KB, kinetic buffer; PBS, phosphate buffered saline; SPR, surface plasmon resonance; antibody recycling; chemokine; histidine; mAb, monoclonal antibody; monoclonal antibody; pH-dependency; phage display; phage libraries; scFv, single-chain variable fragment