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AFFINImeter & Isothermal Titration Calorimetry


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AFFINImeter & Isothermal Titration Calorimetry: A perfect match for the complete Thermodynamic and Kinetic Characterization of molecular interactions

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AFFINImeter & Isothermal Titration Calorimetry

  1. 1. for the complete Thermodynamic and Kinetic Characterization of molecular interactions (1) AFFINImeter Scientific & Development team, Software 4 Science Developments, S. L. Ed. Emprendia. � (2) Dept. of Applied Physics, Fac. of Physics, USC, Campus Vida, Santiago de Compostela, A Coruña 15782, Spain. (3) Biophysics & Structural Biology team, IBMC, UPR9002 du CNRS 15 rue René Descartes, 67084 Strasbourg, France. Website:; E-mail: AFFINImeter & Isothermal Titration Calorimetry 1.- GET THERMODYNAMIC AND KINETIC PROFILES OF YOUR INTERACTION IN A FEW CLICKS AUTOMATIC RAW DATA PROCESSING BASELINE CORRECTION & NOISE ELIMINATION Thermodynamic profile KA = (2.04 ± 0.25)·106 M-1 ∆H = (-9.34 ± 0.13) Kcal·mol-1 * KinITC: AN EXCLUSIVE TOOL TO GET KINETIC INFORMATION THE MODEL BUILDER An easy to use tool to design your own tailored binding models GLOBAL FITTING Perform various experiments and fit globally the resulting isotherms to get the most reliable result A PERFECT MATCH Eva Muñoz1 , Angel Piñeiro1,2 , Juan Sabin1 , Javier Rial1 , Philippe Dumas3 , Eric Ennifar3 2.- GET STRUCTURAL AND MECHANISTIC INFORMATION OF COMPLEX INTERACTING SYSTEMS 3.- QUICK ACCESS TO DATA INTERPRETATION TOOLS PEAK INTEGRATION WITH ASSOCIATED ERRORS Kinetic profile kon = (2.26 ± 0.19)·104 M-1 ·s-1 koff = (1.11 ± 0.09)·10-2 s-1 RAW DATA PROCESSSED DATA ITC ISOTHERM EQUILIBRATION TIME CURVE (ETC) M = RECEPTOR IN CELL A = LIGAND IN SYRINGE B = COMPETING LIGAND COMPETITIVE BINDING OF TWO LIGANDS BINDING TO A THREE SITE RECEPTOR EQUILIBRATION TIME LIGAND “A” INTO RECEPTOR LIGAND “B” INTO RECEPTOR LIGAND “A” INTO (RECEPTOR-LIGAND “B”) Local Minimum Global Minimum FRACTION OF OCCUPIED SITES SITE 1 SITE 2 CONTRIBUTIONS TO THE BINDING ISOTHERM SITE 1 SITE 3 SITE 2 LIGAND INTO MULTIVALENT POLYMER LOCAL MINIMA LIGAND BINDING Nº OF SITES KA (M-1 ) ∆H (Kcal·mol-1 ) Site 1 4-5 (1.93 ± 0.11)·105 -78.14 ± 11.24 Site 2 1-2 (1.68 ± 0.08)·107 -0.14 ± 11.37 Site 3 7-9 (5.30 ± 1.01)·106 -98.44 ± 1.17 SITE 3 References * a) Burnouf D, Ennifar E, Guedich S, Puffer-Enders B, Hoffmann G, Bec G., Disdier F, Baltzinger M, Dumas P (2012) JACS 134, 559-565 b) Dumas P, Ennifar E, Bec G, Piñeiro A, Sabín J, Muñoz E, Rial J, Implementation of KinITC into AFFINImeter, Malvern-MicroCal Application note KinITC: a new method for obtaining joint thermodynamic and kinetic data by isothermal titration calorimetry. (2015).