Advantages of Isothermal Titration Calorimetry: Best to Reveal Differential Binding Thermodynamics
Antigen binding with antibody is a primary step for the successful production of the protective adaptive immune responses. To realize how crucial the evolution of the adaptive immune response is, understanding of the antibody-antigen interactions’ biophysical properties is very much essential. Similar to other protein-to-protein interactions, antibody-antigen complexes arise from the non-covalent interactions together with electrostatic forces, hydrophobic effects and hydrogen bonds. If the topic of discussion goes over analytical techniques, then Isothermal Titration Calorimetry is in the top lists of the selectors. They are often thought as the gold standard specifically for analyzing the intermolecular interactions. The Isothermal Titration Calorimetry also is a followed titrimetiric method based on volumetric technique and ideal for quantitative chemical analysis. In the analysis process, it is ensured that the titrant (a reagent solution) reacts with a titrand solution (an analyte).
How the Titration is performed?
The titration is performed at a constant temperature and pressure. In simple words, a single isothermal titration Calorimetry experiment provides information about the binding enthalpy, the stoichiometry and the equilibrium association constant from which the Gibbs energy and binding entropy can be determined. Therefore, a single isothermal titration Calorimetry experiment provides unswerving access to the valuable thermodynamic potentials allied to the interaction process of enthalpy, entropy and Gibbs energy.
How Isothermal Titration Calorimetry Help Studying The Binding Interactions?
The isothermal titration Calorimetry is obedient to the standardized requirements for an experimental method aiming towards the study of binding interactions. Isothermal titration Calorimetry helps in determining the heat in the process of reaction directly at a constant pressure, relative to the molar enthalpy change connected with that process. If the non-legated state is considered as a reference, then it has been seen that the amount of macromolecule-ligands complex formed is the whole concentration present in the cell. However, there are certain conditions to meet while estimating the binding enthalpy. Those are mentioned below.
· Make sure that complete macromolecular saturation is achieved
· Ensure that ideal subtraction of the reference background heat is performed
· Make certain that protein concentration is found out with rational accuracy
The Way beyond Advantages of Isothermal Titration Calorimetry:
There are many advantages of isothermal titration Calorimetry in comparison to other biophysical techniques. The first ever benefit is it gives an absolute basic thermodynamic characterization in a single experiment. The thermodynamic characterization includes association, stoichiometry and binding enthalpy. The second advantage of using this is it doesn’t require reporter labels such as fluorophores, chromophores etc. as heat is a universal signal. The isothermal titration Calorimetry helps in determining the binding enthalpy directly which is the third advantage of it.
The fourth crucial benefit is the process applies non-destructive technique for production. The fifth benefit of Protein Crystallization is they give better interaction chances in solution. Sixth, they give high possibility to performing experiment with unusual systems or optically dense solutions such as cells or intact organelles, dispersions and so on. The last but not in the least benefit of Isothermal Titration Calorimetry is it is considerably fast during the conventional titration experiment.