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Brunauer, Emmett, and Teller (BET) introduced a theory in 1938 that modified Langmuir's assumptions to explain multilayer physical adsorption. This theory is applicable to Type II isotherms and provides a more realistic picture of adsorption processes. The BET theory assumes a uniform solid surface with localized adsorption sites, where adsorption at one site doesn't affect adsorption at neighboring sites. This theory also allows for the possibility of additional molecules being adsorbed on top...
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Fitting two- and three-site binding models to isothermal titration calorimetric data.

Chad A Brautigam1

  • 1Department of Biophysics, The University of Texas Southwestern Medical Center, Dallas, TX 75390-8816, United States.

Methods (San Diego, Calif.)
|December 9, 2014
PubMed
Summary

Isothermal titration calorimetry (ITC) can analyze complex biomolecular interactions like ABB and ABBB systems. However, data analysis for multi-site binding requires careful examination to ensure accurate thermodynamic parameter estimation.

Keywords:
Analytical ultracentrifugationIsotherm analysisIsothermal titration calorimetryThree-site bindingTreponemal lipoproteinsTwo-site binding

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Area of Science:

  • Biochemistry
  • Biophysical Chemistry
  • Thermodynamics

Background:

  • Isothermal titration calorimetry (ITC) is increasingly used for complex biomolecular interactions beyond simple 1:1 binding.
  • Studying systems with multiple binding sites (e.g., ABB, ABBB) presents challenges in data analysis.
  • The number of injections and data structure significantly impact parameter extraction accuracy.

Purpose of the Study:

  • To assess the capability of empirical and simulated ITC data to support simultaneous thermodynamic parameter estimation for ABB and ABBB systems.
  • To investigate the influence of isotherm shape on the reliability of parameter refinement.
  • To determine the conditions under which ITC data can reliably yield multiple thermodynamic parameters.

Main Methods:

  • Utilized both empirical and simulated isothermal titration calorimetry (ITC) data.
  • Focused on analyzing data from ABB and ABBB binding systems.
  • Evaluated the ability of data to support simultaneous estimation of thermodynamic parameters.

Main Results:

  • Multiphasic isotherms generally support the estimation of multiple thermodynamic parameters, though not always definitively.
  • Uniphasic isotherms from multi-site binding systems pose significant challenges for parameter estimation.
  • The precision of estimated parameters is highly dependent on the specific data characteristics.

Conclusions:

  • The complexity of biomolecular interactions necessitates careful consideration of ITC data structure for accurate thermodynamic analysis.
  • While multiphasic isotherms offer better prospects, thorough data exploration is crucial for all multi-site binding systems.
  • Further investigation into data precision is essential for reliable parameter determination in complex ITC studies.