title: Cross-domain inhibition of TACE ectodomain.
abstract: Proteolytic release from the cell surface is an essential activation event for many growth factors and cytokines. TNF-α converting enzyme (TACE) is a membrane-bound metalloprotease responsible for solubilizing many pathologically significant membrane substrates and is an attractive therapeutic target for the treatment of cancer and arthritis. Prior attempts to antagonize cell-surface TACE activity have focused on small-molecule inhibition of the metalloprotease active site. Given the highly conserved nature of metalloprotease active sites, this paradigm has failed to produce a truly specific TACE inhibitor and continues to obstruct the clinical investigation of TACE activity. We report the bespoke development of a specific TACE inhibitory human antibody using "two-step" phage display. This approach combines calculated selection conditions with antibody variable-domain exchange to direct individual antibody variable domains to desired epitopes. The resulting "cross-domain" human antibody is a previously undescribed selective TACE antagonist and provides a unique alternative to small-molecule metalloprotease inhibition.
authors.affiliation: Department of Oncology, University of Cambridge, Cancer Research UK Cambridge Research Institute, Li Ka Shing Centre, Cambridge CB2 0RE, United Kingdom.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Tape
authors.forename: Sofie H
authors.initials: SH
authors.lastname: Willems
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Dombernowsky
authors.forename: Peter L
authors.initials: PL
authors.lastname: Stanley
authors.forename: Marton
authors.initials: M
authors.lastname: Fogarasi
authors.forename: Willem
authors.initials: W
authors.lastname: Ouwehand
authors.forename: John
authors.initials: J
authors.lastname: McCafferty
authors.forename: Gillian
authors.initials: G
authors.lastname: Murphy
mesh_terms: D051722:ADAM Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000072198:ADAM17 Protein; D000818:Animals; D000906:Antibodies / Q000494:pharmacology*; D015195:Drug Design; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008958:Models, Molecular; D016296:Mutagenesis; D019151:Peptide Library; D017434:Protein Structure, Tertiary / Q000235:genetics; D020349:Surface Plasmon Resonance
chemical_list: D000906:Antibodies; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D051722:ADAM Proteins; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human; C000606463:Adam17 protein, mouse