Utilization of microarrayed compound screening (μARCS) to identify inhibitors of p56lck tyrosine kinase

Protein tyrosine kinases play critical roles in cell signaling and are considered attractive targets for drug discovery. The authors have applied muARCS (microarrayed compound screening) technology to develop a high-throughput screen for finding inhibitors of the p56(lck) tyrosine kinase. Initial assay development was performed in a homogeneous time-resolved (LANCE(TM)) format in 96-well microplates and then converted into the gel-based muARCS format. The muARCS methodology is a well-less screening format in which 8640 compounds are arrayed on a microplate-sized piece of polystyrene and subsequently assayed by placing reagents cast in agarose gels in contact with these compound sheets. A blotting paper soaked with adenosine triphosphate is applied on the gel to initiate the kinase reaction in the gel. Using this screening methodology, 300,000 compounds were screened in less than 40 h. Substantial reagent reduction was achieved by converting this tyrosine kinase assay from a 96-well plate assay to muARCS, resulting in significant cost savings.