A High Content Primary Screening Assay Identifying Multiple Protein Targets in a Cell Using Quantum Dots

Abstract

The development of multiplexing capabilities and high content readouts reporting individual cellular measurements enables assessment of biological variability on a single cell basis. A multiplexed high content screening (HCS) assay allows additional information to be gained from a single assay. One such example is the ability to determine effects of chemical entities on multiple proteins, tested in the same cell.

This approach can greatly increase screening efficiencies and enhance the amount of information achieved from a particular assay. Mitogen-activated protein kinases (MAPKs) are a major target for drug discovery programmes. Using this pathway, we have previously presented data showing the application of HCS to rapidly determine protein kinase activity.

Here, we have used the unique optical properties of semiconductor quantum dots to achieve in-well multiplexing of distinct proteins, including the MAP kinases MEK and ERK. This approach has been applied to rapid high information screening screening approaches in kinase target discovery.