An ACE Method for Monitoring Protein-protein Interactions (PPIs)
Conference Recording Apr 30, 2013
About the SpeakerCarol originally studied Biochemistry at the University of Bath (UK), then finished her studies in London, completing a Welcome funded PhD at the Royal Postgraduate Medical School and a Post-doc at the Imperial Cancer Research Fund. She then joined the James Black Foundation to set up and head the Molecular and Cell Biology Laboratory working on GPCRs. After 7 years, she went to Selcia (Essex, UK) to set up the Biology Department, thus enabling them to offer integrated drug discovery services. Carol has over 20 years’ experience in cell biology, she is currently heading the Biology Group at Selcia and coordinating their fragment screening platform.
Several protein-protein interactions (PPIs) are identified as validated therapeutic targets; however, the challenge with working with this target class is the availability of suitable biophysical techniques to readily detect disruption of protein-protein interactions in solution. Selcia have developed a number of affinity capillary electrophoresis (ACE) assays for PPIs. The technique monitors the electrophoretic mobility of one of the protein partners and any interaction with its protein binding partner in the running buffer is usually observed as a shift in electrophoretic mobility (due to a change in the charge to mass ratio). Inhibition of this mobility shift is observed in the presence of an inhibitor and thus IC50 values can be determined. The technique is microscale, can detect picomolar to millimolar affinities and does not require any protein tethering. Since ACE is a separation technique, it can tolerate protein heterogeneity and is thus ideal for screening natural product extracts.