title: Recommendations for Use and Fit-for-Purpose Validation of Biomarker Multiplex Ligand Binding Assays in Drug Development.
abstract: Multiplex ligand binding assays (LBAs) are increasingly being used to support many stages of drug development. The complexity of multiplex assays creates many unique challenges in comparison to single-plexed assays leading to various adjustments for validation and potentially during sample analysis to accommodate all of the analytes being measured. This often requires a compromise in decision making with respect to choosing final assay conditions and acceptance criteria of some key assay parameters, depending on the intended use of the assay. The critical parameters that are impacted due to the added challenges associated with multiplexing include the minimum required dilution (MRD), quality control samples that span the range of all analytes being measured, quantitative ranges which can be compromised for certain targets, achieving parallelism for all analytes of interest, cross-talk across assays, freeze-thaw stability across analytes, among many others. Thus, these challenges also increase the complexity of validating the performance of the assay for its intended use. This paper describes the challenges encountered with multiplex LBAs, discusses the underlying causes, and provides solutions to help overcome these challenges. Finally, we provide recommendations on how to perform a fit-for-purpose-based validation, emphasizing issues that are unique to multiplex kit assays.
authors.affiliation: Pfizer Inc., One Burtt Road, Andover, Massachusetts, 01810, USA. Darshana.Jani@pfizer.com.
authors.forename: Darshana
authors.initials: D
authors.lastname: Jani
authors.affiliation: LGC Ltd, Newmarket Road, Fordham, Cambridgeshire, CB7 5WW, UK.
authors.forename: John
authors.initials: J
authors.lastname: Allinson
authors.affiliation: Kyowa-Kirin Pharmaceuticals, 212 Carnegie Center #101, Princeton, New Jersey, 08540, USA.
authors.forename: Flora
authors.initials: F
authors.lastname: Berisha
authors.affiliation: Genentech, 1 DNA Way, South San Francisco, California, 94080, USA.
authors.forename: Kyra J
authors.initials: KJ
authors.lastname: Cowan
authors.affiliation: AbbVie Inc., 1 North Waukegan Road North, Chicago, Illinois, 60064, USA.
authors.forename: Viswanath
authors.initials: V
authors.lastname: Devanarayan
authors.affiliation: Bristol-Myers Squibb, Route 206 and Province Line Road, Princeton, New Jersey, 08540, USA.
authors.forename: Carol
authors.initials: C
authors.lastname: Gleason
authors.affiliation: Quanterix Corporation, 113 Hartwell Avenue, Lexington, Massachusetts, 02421, USA.
authors.forename: Andreas
authors.initials: A
authors.lastname: Jeromin
authors.affiliation: Abbvie Inc., 1500 Seaport Blvd, Redwood City, California, 94063, USA.
authors.forename: Steve
authors.initials: S
authors.lastname: Keller
authors.affiliation: KCAS Bioanalytical and Biomarker Services, 12400 Shawnee Mission Parkway, Shawnee, Kansas, 66216, USA.
authors.forename: Masood U
authors.initials: MU
authors.lastname: Khan
authors.affiliation: Radix Biosolutions, 111 Cooperative Way #120, Georgetown, Texas, 78626, USA.
authors.forename: Bill
authors.initials: B
authors.lastname: Nowatzke
authors.affiliation: Quintiles Corporation, 1600 Terrell Mill Road Suite 100, Marietta, Georgia, 30067, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Rhyne
authors.affiliation: Ampersand Biosciences, LLC, 3 Main St., Saranac Lake, New York, 12983, USA.
authors.forename: Laurie
authors.initials: L
authors.lastname: Stephen
mesh_terms: D015415:Biomarkers / Q000032:analysis*; D006801:Humans; D008024:Ligands; D011933:Reagent Kits, Diagnostic / Q000592:standards; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D015415:Biomarkers; D008024:Ligands; D011933:Reagent Kits, Diagnostic
keywords: biomarker; diagnostics; ligand binding assay; multiplex; validation