Development of a IRAK1 Quantitative Protein Degradation Assay for In-vitro and Ex-vivo Target Engagement Using ProteinSimple Capillary Western Technology

IRAK1 is a kinase which has been identified as a key regulator of cytokine signaling and is known to be involved in innate immune responses. Overexpression of these pleotropic cytokines have been implicated in various autoimmune diseases. Antagonists have shown clinical efficacy via modulation of various pro-inflammatory cytokine signals in various mouse models including the collagen induced arthritis (CIA) model. The identified project need was to demonstrate the engagement of IRAK1 with in-house chemical entities in-vitro by generating IC_50s and ex-vivo using mouse tissue from the CIA model. To achieve this goal, a quantitative Capillary Western protein degradation assay was developed.