title: Generation and characterization of a diabody targeting the αvβ6 integrin.
abstract: The αvβ6 integrin is up-regulated in cancer and wound healing but it is not generally expressed in healthy adult tissue. There is increasing evidence that it has a role in cancer progression and will be a useful target for antibody-directed cancer therapies. We report a novel recombinant diabody antibody fragment that targets specifically αvβ6 and blocks its function. The diabody was engineered with a C-terminal hexahistidine tag (His tag), expressed in Pichia pastoris and purified by IMAC. Surface plasmon resonance (SPR) analysis of the purified diabody showed affinity in the nanomolar range. Pre-treatment of αvβ6-expressing cells with the diabody resulted in a reduction of cell migration and adhesion to LAP, demonstrating biological function-blocking activity. After radio-labeling, using the His-tag for site-specific attachment of (99m)Tc, the diabody retained affinity and targeted specifically to αvβ6-expressing tumors in mice bearing isogenic αvβ6 +/- xenografts. Furthermore, the diabody was specifically internalized into αvβ6-expressing cells, indicating warhead targeting potential. Our results indicate that the new αvβ6 diabody has a range of potential applications in imaging, function blocking or targeted delivery/internalization of therapeutic agents.
authors.affiliation: UCL Cancer Institute, University College London, London, United Kingdom.
authors.forename: Heide
authors.initials: H
authors.lastname: Kogelberg
authors.forename: Enrique
authors.initials: E
authors.lastname: Miranda
authors.forename: Jerome
authors.initials: J
authors.lastname: Burnet
authors.forename: David
authors.initials: D
authors.lastname: Ellison
authors.forename: Berend
authors.initials: B
authors.lastname: Tolner
authors.forename: Julie
authors.initials: J
authors.lastname: Foster
authors.forename: Carmen
authors.initials: C
authors.lastname: Picón
authors.forename: Gareth J
authors.initials: GJ
authors.lastname: Thomas
authors.forename: Tim
authors.initials: T
authors.lastname: Meyer
authors.forename: John F
authors.initials: JF
authors.lastname: Marshall
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Mather
authors.forename: Kerry
authors.initials: K
authors.lastname: Chester
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D016023:Integrins / Q000276:immunology* / Q000378:metabolism; D007553:Isotope Labeling; D051379:Mice; D010843:Pichia / Q000235:genetics; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D012878:Skin Neoplasms / Q000378:metabolism; D013667:Technetium / Q000737:chemistry
chemical_list: D000951:Antigens, Neoplasm; D007128:Immunoglobulin Fragments; D016023:Integrins; D011994:Recombinant Proteins; C073878:integrin alphavbeta6; D013667:Technetium
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