title: i-bodies, Human Single Domain Antibodies That Antagonize Chemokine Receptor CXCR4.
abstract: CXCR4 is a G protein-coupled receptor with excellent potential as a therapeutic target for a range of clinical conditions, including stem cell mobilization, cancer prognosis and treatment, fibrosis therapy, and HIV infection. We report here the development of a fully human single-domain antibody-like scaffold termed an "i-body," the engineering of which produces an i-body library possessing a long complementarity determining region binding loop, and the isolation and characterization of a panel of i-bodies with activity against human CXCR4. The CXCR4-specific i-bodies show antagonistic activity in a range of in vitro and in vivo assays, including inhibition of HIV infection, cell migration, and leukocyte recruitment but, importantly, not the mobilization of hematopoietic stem cells. Epitope mapping of the three CXCR4 i-bodies AM3-114, AM4-272, and AM3-523 revealed binding deep in the binding pocket of the receptor.
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083.
authors.forename: Katherine
authors.initials: K
authors.lastname: Griffiths
authors.affiliation: Biomedical Manufacturing, CSIRO Manufacturing, 343 Royal Parade, Parkville, Victoria 3052.
authors.forename: Olan
authors.initials: O
authors.lastname: Dolezal
authors.affiliation: the Australian Regenerative Medicine Institute, Monash University, Wellington Road, Clayton, Victoria 3800,; Biomedical Manufacturing, CSIRO Manufacturing, Bayview Avenue, Clayton, Victoria 3168.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Cao
authors.affiliation: the Australian Regenerative Medicine Institute, Monash University, Wellington Road, Clayton, Victoria 3800,; Biomedical Manufacturing, CSIRO Manufacturing, Bayview Avenue, Clayton, Victoria 3168.
authors.forename: Susan K
authors.initials: SK
authors.lastname: Nilsson
authors.affiliation: the Harry Perkins Institute of Medical Research, Nedlands, Western Australia 6009,; the Centre for Medical Research, University of Western Australia, Crawley, Western Australia 6009.
authors.forename: Heng B
authors.initials: HB
authors.lastname: See
authors.affiliation: the Harry Perkins Institute of Medical Research, Nedlands, Western Australia 6009,; the Centre for Medical Research, University of Western Australia, Crawley, Western Australia 6009,; Dimerix Bioscience Ltd., Nedlands, Western Australia 6009.
authors.forename: Kevin D G
authors.initials: KDG
authors.lastname: Pfleger
authors.affiliation: the Doherty Institute for Infection and Immunity, University of Melbourne, 792 Elizabeth Street, Melbourne, Victoria 3000,; the Burnet Institute, 85 Commercial Road, Melbourne, Victoria 3004.
authors.forename: Michael
authors.initials: M
authors.lastname: Roche
authors.affiliation: the School of Health and Biomedical Sciences, College of Science, Engineering and Health, RMIT University, Melbourne, Victoria 3001.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Gorry
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083.
authors.forename: Andrew
authors.initials: A
authors.lastname: Pow
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083.
authors.forename: Katerina
authors.initials: K
authors.lastname: Viduka
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083.
authors.forename: Kevin
authors.initials: K
authors.lastname: Lim
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083.
authors.forename: Bernadine G C
authors.initials: BGC
authors.lastname: Lu
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083.
authors.forename: Denison H C
authors.initials: DHC
authors.lastname: Chang
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083.
authors.forename: Thomas
authors.initials: T
authors.lastname: Murray-Rust
authors.affiliation: the Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria 3086, and.
authors.forename: Marc
authors.initials: M
authors.lastname: Kvansakul
authors.affiliation: the Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria 3086, and.
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Perugini
authors.affiliation: the Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria 3086, and.
authors.forename: Con
authors.initials: C
authors.lastname: Dogovski
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083.
authors.forename: Marcel
authors.initials: M
authors.lastname: Doerflinger
authors.affiliation: the Department of Medicine, St. Vincent's Hospital, University of Melbourne, Fitzroy, Victoria 3065, Australia.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083,; the Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria 3086, and.
authors.forename: Kathy
authors.initials: K
authors.lastname: Parisi
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083.
authors.forename: Joanne L
authors.initials: JL
authors.lastname: Casey
authors.affiliation: Biomedical Manufacturing, CSIRO Manufacturing, 343 Royal Parade, Parkville, Victoria 3052.
authors.forename: Stewart D
authors.initials: SD
authors.lastname: Nuttall
authors.affiliation: From AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Victoria 3083,; the Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria 3086, and. Electronic address: m.foley@latrobe.edu.au.
authors.forename: Michael
authors.initials: M
authors.lastname: Foley
mesh_terms: D000818:Animals; D000918:Antibody Specificity / Q000276:immunology; D001665:Binding Sites / Q000276:immunology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000276:immunology; D002478:Cells, Cultured; D018360:Crystallography, X-Ray; D018604:Epitope Mapping; D057809:HEK293 Cells; D015658:HIV Infections / Q000276:immunology / Q000517:prevention & control; D018922:HL-60 Cells; D006801:Humans; D019169:Jurkat Cells; D008807:Mice, Inbred BALB C; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D008958:Models, Molecular; D011485:Protein Binding / Q000276:immunology; D000072417:Protein Domains; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors* / Q000276:immunology* / Q000378:metabolism; D061905:Single-Domain Antibodies / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology*; D020349:Surface Plasmon Resonance
chemical_list: D019718:Receptors, CXCR4; D061905:Single-Domain Antibodies
keywords: CXC chemokine receptor type 4 (CXCR-4); G protein-coupled receptor (GPCR); antibody engineering; hematopoietic stem cells; protein structure; single-domain antibody (sdAb, nanobody)