ITC and SPR Analysis Using Dynamic Approach [article]

Janarthanan Krishnamoorthy, Ganesh Kumar Paul Krishanmoorthy, Prashanth Alluvada, Shahul Hameed MS, Russell Pandian, Timothy Kwa
2019 bioRxiv   pre-print
Biophysical techniques such as Isothermal Calorimetry (ITC) and Surface Plasmon Resonance (SPR) are routinely used to ascertain the global binding mechanisms of protein-protein or protein-ligand interaction. Recently, Dominique etal, have explicitly modelled the instrument response of the ligand dilution and analysed the ITC thermogram to obtain kinetic rate constants. Adopting a similar approach, we have integrated the dynamic instrument response with the binding mechanism to simulate the ITC
more » ... rofiles of equivalent and independent binding sites, equivalent and sequential binding sites and aggregating systems. The results were benchmarked against the standard commercial software Origin-ITC. Further, the experimental ITC chromatograms of 2-CMP + RNASE and BH3I-1 + hBCLXL interactions were analysed and shown to be comparable with that of the conventional analysis. Dynamic approach was applied to simulate the SPR profiles of a two-state model, and could reproduce the experimental profile accurately.
doi:10.1101/788075 fatcat:d5kjuedvq5beppvyxukpgik73u