ArticleBiochimica et biophysica acta. General subjects2017
Evaluation and Minimization of Uncertainty in ITC Binding Measurements: Heat Error, Concentration Error, Saturation, and Stoichiometry.
Article in Biochimica et biophysica acta. General subjects, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- Applications of Isothermal Titration Calorimetry in Studying Biomimetic Nanocarriers.Biomolecules · 2025Review
- Modelling the Decamerisation Cycle of PRDX1 and the Inhibition-like Effect on Its Peroxidase Activity.Antioxidants (Basel, Switzerland) · 2023Article
- Nano-Sized Fucoidan Interpolyelectrolyte Complexes: Recent Advances in Design and Prospects for Biomedical Applications.International journal of molecular sciences · 2023Review
- Isothermal Titration Calorimetry for Quantification of Protein-Carbohydrate Interactions.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Structural basis for anti-CRISPR repression mediated by bacterial operon proteins Aca1 and Aca2.The Journal of biological chemistry · 2021Article
- Isothermal titration calorimetry (ITC): a standard operating procedure (SOP).European biophysics journal : EBJ · 2021Article
- A multi-laboratory benchmark study of isothermal titration calorimetry (ITC) using CaEuropean biophysics journal : EBJ · 2021Article
- TcpC inhibits toll-like receptor signaling pathway by serving as an E3 ubiquitin ligase that promotes degradation of myeloid differentiation factor 88.PLoS pathogens · 2021Article
- Experimental characterization of the association of β-cyclodextrin and eight novel cyclodextrin derivatives with two guest compounds.Journal of computer-aided molecular design · 2021Article
- Isothermal titration calorimetry and surface plasmon resonance analysis using the dynamic approach.Biochemistry and biophysics reports · 2020Article
- Estimation of non-constant variance in isothermal titration calorimetry using an ITC measurement model.PloS one · 2020Article
- Obtaining precise and accurate results by ITC.European biophysics journal : EBJ · 2019Article
- vWA proteins of Leptospira interrogans induce hemorrhage in leptospirosis by competitive inhibition of vWF/GPIb-mediated platelet aggregation.EBioMedicine · 2018Article
- A novel Fas-binding outer membrane protein and lipopolysaccharide of Leptospira interrogans induce macrophage apoptosis through the Fas/FasL-caspase-8/-3 pathway.Emerging microbes & infections · 2018Article
- Insights into the interaction mechanism between tiagabine hydrochloride and two serum albumins.RSC advances · 2018Article
- Accounting for apparent deviations between calorimetric and van't Hoff enthalpies.Biochimica et biophysica acta. General subjects · 2018Article
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3 authors.
Funding
Abstract
backgroundIsothermal titration calorimetry (ITC) is uniquely useful for characterizing binding thermodynamics, because it straightforwardly provides both the binding enthalpy and free energy. However, the precision of the results depends on the experimental setup and how thermodynamic results are obtained from the raw data.
methodsExperiments and Monte Carlo analysis are used to study how uncertainties in injection heat and concentration propagate to binding enthalpies in various scenarios. We identify regimes in which it is preferable to fix the stoichiometry parameter, N, and evaluate the reliability of uncertainties provided by the least squares method.
resultsThe noise in the injection heat is mainly proportional in character, with ~1% and ~3% uncertainty at 27C and 65C, respectively; concentration errors are ~1%. Simulations of experiments based on these uncertainties delineate how experimental design and curve fitting methods influence the uncertainty in the final results.
conclusionsIn most cases, experimental uncertainty is minimized by using more injections and by fixing N at its known value. With appropriate technique, the uncertainty in measured binding enthalpies can be kept below ~2% under many conditions, including low C values. GENERAL SIGNIFICANCE: We quantify uncertainties in ITC data due to heat and concentration error, and identify practices to minimize these uncertainties. The resulting guidelines are important when ITC data are used quantitatively, such as to test computer simulations of binding. Reproducibility and further study are supported by free distribution of the new software developed here.
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