Homogeneous Assay for Target Engagement Utilizing Bioluminescent Thermal Shift.

ABSTRACT: Protein thermal shift assays (TSAs) provide a means for characterizing target engagement through ligand-induced thermal stabilization. Although these assays are widely utilized for screening libraries and validating hits in drug discovery programs, they can impose encumbering operational requirements, such as the availability of purified proteins or selective antibodies. Appending the target protein with a small luciferase (NanoLuc) allows coupling of thermal denaturation with luminescent output, providing a rapid and sensitive means for assessing target engagement in compositionally complex environments such as permeabilized cells. The intrinsic thermal stability of NanoLuc is greater than mammalian proteins, and our results indicate that the appended luciferase does not alter thermal den

SUBMITTER: Dart ML 

PROVIDER: S-EPMC6004564 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature