Abstract

High throughput screening (HTS) technologies play an essential role in drug discovery and development. Combining HTS with multiplexing is valuable for accelerating screening processes, but technically challenging and preventatively expensive due to the high cost of required committed instrumentation for closed multiplex systems. Here we describe a new assay platform - FirePlex-384®, which employs data high content imaging (HCI), HTS and multiplexing to harvest data at an unprecedented scale, informing better decisions and accelerating drug discovery. Utilizing patented FirePlex hydrogel particles and a three-region encoding design, FirePlex Immunoassays enable true, in-well multiplexing, providing flexible and customizable quantitation of analytes using established HCI instrumentation. FirePlex®-384 immunoassays quantitate up to 10 protein analytes per well from low sample inputs, in 384-well plate format. The optional two-step workflow or no-wash assay formats limit hands-on time and are compatible with high-throughput automation. Assay data-acquisition is conducted on standard high-content imagers, and data analysis is achieved with the FirePlex® Analysis Workbench software, eliminating the need for dedicated instrumentation and expensive software licenses. By generating more actionable data during early compound screening, FirePlex-384 promptly identifies compounds with the desired activity profile and guides the elimination of compounds with off-target effects early on, thus reducing the number of compounds that will move forward to expensive secondary screens.