2 research outputs found
The Dutch beggars
Thesis (B.L.)--University of Illinois, 1882.Ms.Bound with 8 other humanities theses. IU-
Determination of Ligand-Binding Affinity (<i>K</i><sub>d</sub>) Using Transverse Relaxation Rate (<i>R</i><sub>2</sub>) in the Ligand-Observed <sup>1</sup>H NMR Experiment and Applications to Fragment-Based Drug Discovery
High hit rates from initial ligand-observed NMR screening
can make
it challenging to prioritize which hits to follow up, especially in
cases where there are no available crystal structures of these hits
bound to the target proteins or other strategies to provide affinity
ranking. Here, we report a reproducible, accurate, and versatile quantitative
ligand-observed NMR assay, which can determine Kd values of fragments in the affinity range of low μM
to low mM using transverse relaxation rate R2 as the observable parameter. In this study, we examined the
theory and proposed a mathematical formulation to obtain Kd values using non-linear regression analysis. We designed
an assay format with automated sample preparation and simplified data
analysis. Using tool compounds, we explored the assay reproducibility,
accuracy, and detection limits. Finally, we used this assay to triage
fragment hits, yielded from fragment screening against the CRBN/DDB1
complex