Proximity ligation assays for Hit Profiling in small molecule drug discovery campaigns – a case stud

Proximity ligation is a highly specific and sensitive technology to monitor individual proteins, their interactions and post-translational modifications in situ. It provides single molecule resolution and objective and unbiased quantification in cells and tissues on endogenous expression levels. The European ScreeningPort has optimized the PLA® technology for use in an automated 384-well screening format and has also implemented routines for automated image analysis including spot detection, localisation and quantification. In this presentation, we will present a case study of a small molecule drug discovery project performed in collaboration with a biotech partner. The target mechanism involved the disruption of the interaction between amyloid ß oligomers and RAGE (Receptor for Advanced Glycation Endproducts). Profiling of compounds during Hit-to-Lead stages has involved a variety of High Content functional cellular assays including PLA. We will discuss the use of PLA in 384-well format as part of a Hit-to-Lead screening cascade to monitor compound-mediated disruption of this receptor-ligand complex.