biostudies-literatureUnknown5(6)Finger JNTherapies that suppress RIPK1 kinase activity are emerging as promising therapeutic agents for the treatment of multiple inflammatory disorders. The ability to directly measure drug binding of a RIPK1 inhibitor to its target is critical for providing insight into pharmacokinetics, pharmacodynamics, safety and clinical efficacy, especially for a first-in-class small-molecule inhibitor where the mechanism has yet to be explored. Here, we report a novel method for measuring drug binding to RIPK1 protein in cells and tissues. This TEAR1 (Target Engagement Assessment for RIPK1) assay is a pair of immunoassays developed on the principle of competition, whereby a first molecule (ie, drug) prevents the binding of a second molecule (ie, antibody) to the target protein. Using the TEAR1 assay, we havPharmacology research & perspectiveshttps://www.ebi.ac.uk/biostudies/studies/S-EPMC5723705biostudies-literatureIdentification of an antibody-based immunoassay for measuring direct target binding of RIPK1 inhibitors in cells and tissues.PMC5723705Maguire SNagilla RCampobasso NCook MNRivera EJBrusq JMGough PJHaag HBertin JJoglekar DDeutsch JLich JDHarris PAJenkins ELVotta BJSun HFinger JNfalseIdentification of an antibody-based immunoassay for measuring direct target binding of RIPK1 inhibitors in cells and tissues.Therapies that suppress RIPK1 kinase activity are emerging as promising therapeutic agents for the treatment of multiple inflammatory disorders. The ability to directly measure drug binding of a RIPK1 inhibitor to its target is critical for providing insight into pharmacokinetics, pharmacodynamics, safety and clinical efficacy, especially for a first-in-class small-molecule inhibitor where the mechanism has yet to be explored. Here, we report a novel method for measuring drug binding to RIPK1 protein in cells and tissues. This TEAR1 (Target Engagement Assessment for RIPK1) assay is a pair of immunoassays developed on the principle of competition, whereby a first molecule (ie, drug) prevents the binding of a second molecule (ie, antibody) to the target protein. Using the TEAR1 assay, we hav2017-01-01T00:00:00Z2017 Dec2025-04-21T16:17:22.881Z2019-03-27T03:04:22ZS-EPMC57237052922662510.1002/prp2.377