Rapid and cost effective immunoassay development and fit-for-purpose validation of a biomarker assay using Spatial Proximity Analyte Reagent Capture Luminescence (SPARCL) (TECH2P.760)

Abstract A novel detection technology was used for rapid immunoassay development and validation. SPARCL (Spatial Proximity Analyte Reagent Capture Luminescence) is a proximity dependent and homogeneous chemiluminescent detection method. In a SPARCL assay, a chemiluminescent substrate (acridan) is brought into the proximity of an oxidative enzyme (horseradish peroxidase) through the specific antigen/antibody interaction. A flash of light is generated and is proportional to the amount of IL-8 bound. Assay development was completed in less than 3 days. The validated assay features a wide dynamic range (2 - 4,000 pg/mL), low MRD, short assay run time (60 min.), and sensitivity (2 pg/mL). Quality control samples (QC’s) run in plasma resulted in statistics that show the assay is precise and accurate, with a total error of all QC levels of under 12%. Matrix spiked with IL-8 diluted in a linear manor. The SPARCL technology enables rapid assay development, delivers an assay with desirable performance characteristics and allows for considerable savings in labor, disposables and capital equipment compared to popular platforms. Existing software and luminometers may be used for a SPARCL data acquisition. As a result, there may not be a need to validate new software or plate readers already running in existing Laboratory Information Management Systems or in a regulated environment. The SPARCL assay is microtiter plate-based and is suitable for use with robotics and in HTS applications.