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P2025
High-Density STEP Array Plates  A Novel Platform for Multiplexed GPCR Screening
Presenter Maria Ines Morano, Originus, Inc., USA
Additional Authors: Chi S. Ho, Adam W. Avery
High-density STEP array plates, a novel application of Surface Transfection and Expression Protocol (STEP) technology, have the potential to dramatically increase the high-throughput capacity of cell-based assays used in drug screening. A high-density STEP array plate is generated by printing in each microplate well an array of STEP complex spots containing distinct DNAs. Upon addition by the end user of cells to the microplate, an ordered array of transiently transfected cell populations is obtained within each well. Typical cell-based assays used in drug screening allow a single test compound to be tested against one GPCR target per well. However, high density STEP array plates offer the ability to simultaneously test a single compound against multiple different GPCR targets in a single well.
We present the application of STEP technology to generate high-density STEP array plates for a multiplexed assay for GPCRs related to feeding behavior. We demonstrate the ability to generate ordered groups of distinct transiently transfected populations within microplate wells using HEK293T cells or neural cell lines. Proprietary protein components of STEP complex along with optimized microplate well surface chemistry ensure excellent STEP complex spot and cell retention at the spot location. Typically each spot of STEP complex yields over five-hundred transfected cells, and multiple plasmids can be efficiently co-transfected from a single STEP complex spot. Further, we demonstrate that the stoichiometry of co-transfected plasmids can be controlled, a feature important for development of a cell-based assay for drug screening.
We show the successful application of STEP arrays for simultaneously measuring cellular responses from distinct cell populations within a single well. This demonstrates utility of STEP arrays as a platform for multiplexed GPCR assays in primary or secondary drug screening.