Tags

Type your tag names separated by a space and hit enter

Dual display: phage selection driven by co-engagement of two targets by two different antibody fragments.
Protein Eng Des Sel 2017; 30(9):575-582PE

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.

Authors+Show Affiliations

Department of Pathology and Immunology, University of Geneva, Geneva, CH-1211, Switzerland. Selexis SA, Plan-les-Ouates, CH-1228, Switzerland.Department of Pathology and Immunology, University of Geneva, Geneva, CH-1211, Switzerland.Department of Pathology and Immunology, University of Geneva, Geneva, CH-1211, Switzerland.Department of Pathology and Immunology, University of Geneva, Geneva, CH-1211, Switzerland.NovImmune SA, Plan-les-Ouates, CH-1228, Switzerland.NovImmune SA, Plan-les-Ouates, CH-1228, Switzerland.NovImmune SA, Plan-les-Ouates, CH-1228, Switzerland. Glenmark Pharmaceuticals SA, La Chaux de Fonds, CH-2300 Switzerland.NovImmune SA, Plan-les-Ouates, CH-1228, Switzerland.NovImmune SA, Plan-les-Ouates, CH-1228, Switzerland.Department of Pathology and Immunology, University of Geneva, Geneva, CH-1211, Switzerland.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

28444391

Citation

Fagète, Séverine, et al. "Dual Display: Phage Selection Driven By Co-engagement of Two Targets By Two Different Antibody Fragments." Protein Engineering, Design & Selection : PEDS, vol. 30, no. 9, 2017, pp. 575-582.
Fagète S, Botas-Perez L, Rossito-Borlat I, et al. Dual display: phage selection driven by co-engagement of two targets by two different antibody fragments. Protein Eng Des Sel. 2017;30(9):575-582.
Fagète, S., Botas-Perez, L., Rossito-Borlat, I., Adea, K., Gueneau, F., Ravn, U., ... Hartley, O. (2017). Dual display: phage selection driven by co-engagement of two targets by two different antibody fragments. Protein Engineering, Design & Selection : PEDS, 30(9), pp. 575-582. doi:10.1093/protein/gzx021.
Fagète S, et al. Dual Display: Phage Selection Driven By Co-engagement of Two Targets By Two Different Antibody Fragments. Protein Eng Des Sel. 2017 09 1;30(9):575-582. PubMed PMID: 28444391.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Dual display: phage selection driven by co-engagement of two targets by two different antibody fragments. AU - Fagète,Séverine, AU - Botas-Perez,Ledicia, AU - Rossito-Borlat,Irène, AU - Adea,Kenneth, AU - Gueneau,Franck, AU - Ravn,Ulla, AU - Rousseau,François, AU - Kosco-Vilbois,Marie, AU - Fischer,Nicolas, AU - Hartley,Oliver, PY - 2016/10/07/received PY - 2017/04/13/accepted PY - 2017/4/27/pubmed PY - 2017/11/29/medline PY - 2017/4/27/entrez KW - antibody KW - bispecific KW - leucine zipper KW - phage display SP - 575 EP - 582 JF - Protein engineering, design & selection : PEDS JO - Protein Eng. Des. Sel. VL - 30 IS - 9 N2 - 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. SN - 1741-0134 UR - https://www.unboundmedicine.com/medline/citation/28444391/Dual_display:_phage_selection_driven_by_co_engagement_of_two_targets_by_two_different_antibody_fragments_ L2 - https://academic.oup.com/peds/article-lookup/doi/10.1093/protein/gzx021 DB - PRIME DP - Unbound Medicine ER -