RT Journal Article
SR Electronic
T1 Avoiding accuracy-limiting pitfalls in the study of protein-ligand interactions with isothermal titration calorimetry
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 023796
DO 10.1101/023796
A1 Sarah E. Boyce
A1 Joel Tellinghuisen
A1 John D. Chodera
YR 2015
UL http://biorxiv.org/content/early/2015/08/03/023796.abstract
AB Isothermal titration calorimetry (ITC) can yield precise (3%) estimates of the thermodynamic parameters describing biomolecular association (affinity, enthalpy, and entropy), making it an indispensable tool for biochemistry and drug discovery. Surprisingly, interlaboratory comparisons suggest that errors of ∽ 20% are common and widely underreported. Here, we show how to reduce precision- and accuracy-limiting errors while obtaining good estimates and minimizing material and time consumed by an experiment. We provide a simple spreadsheet that allows practitioners to identify precision-limiting operations during protocol design, track precision during the experiment, and propagate error to yield realistic final uncertainties.