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Dual display: phage selection driven by co-engagement of two targets by two different antibody fragments

Gueneau, Franck
Ravn, Ulla
Rousseau, François
Kosco-Vilbois, Marie
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Published in Protein engineering, design & selection. 2017, vol. 30, no. 9, p. 575-582
Abstract Antibody phage display technology has supported the emergence of numerous therapeutic antibodies. The development of bispecific antibodies, a promising new frontier in antibody therapy, could be facilitated by new phage display approaches that enable pairs of antibodies to be co-selected based on co-engagement of their respective targets. We describe such an approach, making use of two complementary leucine zipper domains that heterodimerize with high affinity. Phagemids encoding a first antibody fragment (scFv) fused to phage coat protein via the first leucine zipper are rescued in bacteria expressing a second scFv fused to the second leucine zipper as a soluble periplasmic protein, so that it is acquired by phage during assembly. Using a soluble scFv specific for a human CD3-derived peptide, we show that its acquisition by phage displaying an irrelevant antibody is sufficiently robust to drive selection of rare phage (1 in 10(5)) over three rounds of panning. We then set up a model selection experiment using a cell line expressing the chemokine receptor CCR5 fused to the CD3 peptide together with a panel of phage clones capable displaying either an anti-CCR5 scFv or an irrelevant antibody, with or without the capacity to acquire the soluble anti-CD3 scFv. In this experiment we showed that rare phage (1 in 10(5)) capable of displaying the two different scFvs can be specifically enriched over four rounds of panning. This approach has the potential to be applied to the identification of pairs of ligands capable of co-engaging two different user-defined targets, which would facilitate the discovery of novel bispecific antibodies.
Keywords Amino Acid SequenceAnimalsAntibodiesBispecific/biosynthesis/geneticsAntibodiesMonoclonal/biosynthesis/geneticsAntibody SpecificityBacteriophages/genetics/immunologyCD3 Complex/genetics/immunologyCHO CellsCapsid Proteins/genetics/immunologyCricetulusEscherichia coli/genetics/metabolism/virologyGene ExpressionHumansLeucine ZippersPeptide LibraryPeptides/genetics/immunologyReceptorsCCR5/genetics/immunologyRecombinant Fusion Proteins/genetics/immunologySequence AnalysisDNASingle-Chain Antibodies/biosynthesis/genetics
PMID: 28444391
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Research group Hiv (835)
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FAGETE, Séverine et al. Dual display: phage selection driven by co-engagement of two targets by two different antibody fragments. In: Protein Engineering, Design and Selection, 2017, vol. 30, n° 9, p. 575-582. doi: 10.1093/protein/gzx021 https://archive-ouverte.unige.ch/unige:112661

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Deposited on : 2019-01-09

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