Fecha registrada: May 21 2015

Duration: 30 minutes 34 seconds

The primary signal measured by a microcalorimeter is a heat power; that is the rate of production of the heat evolved during a reaction. As such, this primary signal is directly linked to the kinetics of the reaction. kinITC has been developed to recover this kinetic information from the shape of each injection curve in a classical ITC experiment. It will be shown how the true kinetic signal is affected by the response time of the instrument. In situations where the reaction (i) is not too rapid and (ii) follows a one-step mechanism, the kinetic parameters kon and koff can be recovered immediately after the regular treatment yielding the △H and the KD of the reaction. This has been implemented in the software AFFINIMeter.

Table of contents
1. Welcome
02:22
2. kinITC: Get More Out of ITC Data
01:28
3. Why is ITC also a Kinetic Technique
01:07
4. ITC as a tool...
00:50
5. Why is ITC also a kinetic technique?
00:05
6. Integration of each injection
00:49
7. There is kinetic information embedded in the shape of the injection curves
00:47
8. Examples
01:10
9. The basics of kinITC
00:04
10. How retrieving this kinetic information?How transforming ITC into kinITC for obtaining thermodynamic and kinetic information in general situations ?
00:19
11. Let us consider the simplest situation One single kinetic step
02:12
12. Simulation of one injection
01:01
13. Simulation of an ITC experiment
01:24
14. The practical problems
00:06
15. The practical problemsParticularly for fast kinetics
00:38
16. How to measure the instrument response time?
00:48
17. How to measure the instrument response time?
00:28
18. Exponential fit of each return to baseline
01:12
19. Influence of the instrument response time
01:22
20. Is there kinetic information in my data?
00:14
21. Is there a variation of the equilibration time?
00:43
22. Simple consequence: compare the titration curve with the envelope of the injection curves
01:54
23. AFFINImeter
00:40
24. The simplified kinITC method: kon & koff with one click in the software AFFINImeter
00:20
25. Evolution of the equilibration time during a titration experiment
00:25
26. Evolution of the equilibration time during a titration experiment
00:48
27. Illustration with a simple case Carbonic anhydrase + inhibitor (4CBS)
00:25
28. Results for carbonic anhydrase at several temperatures
00:52
29. ETC-based kinITC
00:35
30. ETC-based kinITC
00:22
31. AFFINImeter
00:22
32. Comparison of results with SPR
01:23
33. Comparison of results with SPR
01:25
34. Take-home message
00:34
35. Contact Information
01:20
36. For further information on kinITC, please see the following resources:
00:00