Disentangling the origin of chemical differences using GHOST
Affinimeter Tutorial
1. AFFINImeter: una herramienta avanzada
para el análisis de datos de
Calorimetría de Titulación Isotérmica (ITC)
TUTORIAL
15 y 16 de Octubre de 2015
2. MOLECULAR RECOGNITION
Essential for all physiological and pathological processes.
• Cell grouth and development
• Cell-cell communication
• Bacterial/viral infections
• Tumor development
• etc…..
3. MOLECULAR RECOGNITION
Ligand – receptor interactions
• Protein – protein
• Nucleic acid – protein
• Small molecule – biomolecule…….
Free species Complex
A + M AM
4. o Understanding Biological proceses.
o Pathological proceses.
o Drug Discovery and development.
Ligand – receptor interactions: characterization
• Thermodynamics
• Kinetics
• Structure
• How strong?
• How fast?
• Where/how?
8. Thermodynamics
• Gold-standard technique for the thermodynamic characterization of interactions
• Measures the heat change occured in the calorimetric cell throughout the titration
ligand
receptor
The ITC titration:
Power vs. Time plot (thermogram)
ITC isotherm
peak integration
KA , DH, stoichiometry
DG, DS
12. Obtaining kinetic information from ITC
• IS THERE KINETIC INFORMATION IN MY THERMOGRAM?: A VISUAL INSPECTION
Mid titration: Peak broadening
KinITC: obtaining kinetic information from ITC
14. “Essentially, KinITC is based upon linking the kinetics of the reaction to heat power
production in the measurement cell”
=
=
= = concentration of titrand in the cell
1:1 binding
Heat power
KinITC: obtaining kinetic information from ITC
AFFINImeter & KinITC
15. Wieseman parameter
Stoichiometric ratio for current injection
KinITC: obtaining kinetic information from ITC
Equilibration time
Dissociation rate constant
AFFINImeter & KinITC