High-Throughput Screening Applications of a Novel Scanning Laser Imaging Technology

Presented at the October 7^th, 1999 meeting of the LRIG Mid Atlantic

The increasing number of compounds available for screening in drug development has driven the requirement for higher throughput screening technologies, as well as unique technologies that address a broader application portfolio. Our laboratory has been involved in the development of multiplexed mix-and-read assays using Fluorometric Microvolume Assay Technology (FMAT). FMAT is a fluorescence based assay system that incorporates a laser scanner and optical detection system that provides a direct measurement of cellular or bead-based fluorescence on a well-to-well and on an individual cell/bead basis. This design is ideal for the homogenous identification of hits in primary screening, as well as for lead optimization in the form of IC50 determinations, and for assessment of lead compound cytotoxicity. Fluorescent beads of various sizes can be distinguished from one another, allowing the multiplexing of two or more targets present on different sized beads in the same well. In addition, the digitized image data is compiled from two PMTs permitting the development of multiplexed assays based on dye color. A variety of different mix and read applications for FMAT will be described, including peptide-receptor ligand interactions, and multiplexed bead based immunocapture assays. The results of a novel high-throughput screen performed in collaboration with a major Pharma oncology group will also be discussed. Employing a simple mix-and-read Annexin V binding assay, a variety of purified natural products were identified that are potent inducers of apoptosis in tumor cells. Taken together, the data to be presented will demonstrate the versatility and feasibility of fluorescence-based homogeneous and multiplexed assays for a variety of cell-based and molecular targeted screens used in drug discovery.

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