Easily Automate PK Assays for Human IgG Drugs in Preclinical Species
News Jul 27, 2017
Image Credit: Andrew Alliance
Lumigen Inc. (A Beckman Coulter Company) presented a poster demonstrating the use of Andrew the pipetting robot with their SPARCL Pharmacokinetic Immunoassay kits at the AAPS NBC event in San Diego. The poster summarizes Andrew’s accuracy and error free execution of the SPARCL kits leading to improved efficiency, reduced PK assay development and cost, all while increasing profitability. The specific poster relates to measuring IgG drug concentration in a pre-clinical setting.
The combination of these two unique technologies allows for a typical ELISA workflow that takes days from the start of the process (starting with coating plates) into just a couple of hours from start to finish.
The data and resulting analysis concludes that the combination of the Andrew robot with the Life Diagnostics SPARCL PK kit may be a way for CRO’s to improve efficiencies, reduce PK assay development time and cost, reduce labor and increase profitability.
SPARCLTM (Spatial Proximity Analyte Reagent Capture Luminescence) technology is a proximity dependent chemiluminescent detection method. In a SPARCL assay, a chemiluminescent compound (acridan) is brought into close proximity to an oxidative enzyme (horseradish peroxidase, HRP) through specific antigen/antibody interaction. A flash of light proportional to the quantity of analyte present in the sample is generated upon addition of a trigger solution containing hydrogen peroxide. A background reducing agent (BGR) is often used to enhance S/N ratios. There is no need to remove excess reactants. There are no washing steps.
The SPARCL PK kit was run according to the directions provided by the manufacturer (Life Diagnostics) by giving the pipetting instructions to the Andrew robot. Briefly, Andrew made the dilutions for the standard curve and QC’s, pipetted the antibodies, calibrators and QC’s into the wells of the 96 well plates. The 96 well plate was incubated at room temperature for 30 minutes on a orbital shaker set at 350 revolutions per minute. After the incubation time was complete, the plate was moved to the LUMIstar Omega luminometer. The LUMIstar was instructed to inject the trigger solution and collect the emitted light.