title: Recommendations for the development and validation of flow cytometry-based receptor occupancy assays.
abstract: Receptor occupancy measurements demonstrate the binding of a biotherapeutic agent to its extra-cellular target and represent an integral component of the pharmacodynamic (PD) portfolio utilized to advance the development and commercialization of a therapeutic agent. Coupled with traditional pharmacokinetic (PK) assessments derived from serum drug concentration, receptor occupancy data can be used to model PK/PD relationships and validate dose selection decisions throughout the drug development lifecycle. Receptor occupancy assays can be even more challenging to develop than other flow cytometric methods (e.g. surface immunophenotyping). In addition to typical considerations regarding stability of the cell type of interest, stability of the target-bound therapeutic agent and stability of the target receptor must be taken into account. Reagent selection is also challenging as reagents need to be evaluated for the potential to compete with the therapeutic agent and bind with comparable affinity. This article provides technical guidance for the development and validation of cytometry-based receptor occupancy assays.
authors.affiliation: Amgen, Inc, 1 Amgen Center Drive, Mailstop 30E-3-C, Thousand Oaks, California, 91320.
authors.forename: Cherie L
authors.initials: CL
authors.lastname: Green
authors.affiliation: Flow Contract Site Laboratory, LLC, 13029 NE 126th PL, Unit A229, Kirkland, Washington, 98034.
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Stewart
authors.affiliation: Novo Nordisk a/S, Novo Nordisk Park, Måløv, DK, 2760, Denmark.
authors.forename: Carl-Magnus
authors.initials: CM
authors.lastname: Högerkorp
authors.affiliation: Laboratory Corporation of America® Holdings, LabCorp Clinical Trials, 201 Summit View Dr, Suite 200, Brentwood, Tennessee, 37027.
authors.forename: Alan
authors.initials: A
authors.lastname: Lackey
authors.affiliation: Laboratory Corporation of America® Holdings, LabCorp Clinical Trials, 201 Summit View Dr, Suite 200, Brentwood, Tennessee, 37027.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Jones
authors.affiliation: Medimmune, LLC, 319 North Bernardo Avenue, Mountain View, California, 94043.
authors.forename: Meina
authors.initials: M
authors.lastname: Liang
authors.affiliation: Alnylam Pharmaceuticals, Bioanalytical Sciences, 300 Third Street, Cambridge, Massachusetts, 02142.
authors.forename: Yuanxin
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Amgen, Inc, 1 Amgen Center Drive, Mailstop 30E-3-C, Thousand Oaks, California, 91320.
authors.forename: John
authors.initials: J
authors.lastname: Ferbas
authors.affiliation: BioCytex, 140 Chemin De L'armée D'afrique, Marseille, 13010, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Moulard
authors.affiliation: 7 Rue Moïse-Marcinhes, Geneva, Meyrin, 1217, Switzerland.
authors.forename: Kamila
authors.initials: K
authors.lastname: Czechowska
authors.affiliation: ICON Laboratory Services, 123 Smith Street, Farmingdale, New York, 11735.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Mc Closkey
authors.affiliation: PRA Healthsciences, Amerikaweg 18, Assen, TK, 9407, The Netherlands.
authors.forename: Barry W A
authors.initials: BW
authors.lastname: van der Strate
authors.affiliation: Charles River Laboratories International, Inc, 6995 Longley Lane, Reno, Nevada, 89511.
authors.forename: Danice E C
authors.initials: DE
authors.lastname: Wilkins
authors.affiliation: Eurofins Pharma Bioanalysis Services UK Limited, 91 Park Drive, Milton Park Abingdon, OX14 4RY, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Lanham
authors.affiliation: Takeda Pharmaceuticals, 35 Landsdown St, Cambridge, Massachusetts, 02139.
authors.forename: Timothy
authors.initials: T
authors.lastname: Wyant
authors.affiliation: Covance Central Laboratory Services, 8211 SciCor Dr, Indianapolis, Indiana, 46214.
authors.forename: Virginia
authors.initials: V
authors.lastname: Litwin
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000627:therapeutic use; D055808:Drug Discovery; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000627:therapeutic use; D006801:Humans
chemical_list: D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes
keywords: biotherapeutic; flow cytometry; pharmacodynamic biomarker; pharmacokinetics; receptor occupancy; target binding