title: Probing a protein-protein interaction by in vitro evolution.
abstract: In this study, we used in vitro protein evolution with ribosome and phage display to optimize the affinity of a human IL-13-neutralizing antibody, a therapeutic candidate for the treatment of asthma, >150-fold to 81 pM by using affinity-driven stringency selections. Simultaneously, the antibody potency to inhibit IL-13-dependent proliferation in a cell-based functional assay increased 345-fold to an IC50 of 229 pM. The panoply of different optimized sequences resulting from complementarity-determining region-targeted mutagenesis and error-prone PCR using ribosome display was contrasted with that of complementarity-determining region-targeted mutagenesis alone using phage display. The data highlight the advantage of the ribosome-display approach in identifying beneficial mutations across the entire sequence space. A comparison of mutation hotspots from in vitro protein evolution to knockout mutations from alanine scanning demonstrated that in vitro evolution selects the most appropriate positions for improvements in potency without mutating any of the key residues within the functional paratope.
authors: Cambridge Antibody Technology, Milstein Building, Granta Park, Cambridge CB1 6GH, United Kingdom.
authors: George
authors: G
authors: Thom
authors: Alexis C
authors: AC
authors: Cockroft
authors: Andrew G
authors: AG
authors: Buchanan
authors: Cathy Joberty
authors: CJ
authors: Candotti
authors: E Suzanne
authors: ES
authors: Cohen
authors: David
authors: D
authors: Lowne
authors: Phill
authors: P
authors: Monk
authors: Celia P
authors: CP
authors: Shorrock-Hart
authors: Lutz
authors: L
authors: Jermutus
authors: Ralph R
authors: RR
authors: Minter