| 00:00:00 | Welcome |
| 00:02:22 | kinITC: Get More Out of ITC Data |
| 00:03:50 | Why is ITC also a Kinetic Technique |
| 00:04:57 | ITC as a tool... |
| 00:05:47 | Why is ITC also a kinetic technique? |
| 00:05:52 | Integration of each injection |
| 00:06:41 | There is kinetic information embedded in the shape of the injection curves |
| 00:07:28 | Examples |
| 00:08:38 | The basics of kinITC |
| 00:08:42 | How retrieving this kinetic information?How transforming ITC into kinITC for obtaining thermodynamic and kinetic information in general situations ? |
| 00:09:01 | Let us consider the simplest situation One single kinetic step |
| 00:11:13 | Simulation of one injection |
| 00:12:14 | Simulation of an ITC experiment |
| 00:13:38 | The practical problems |
| 00:13:44 | The practical problemsParticularly for fast kinetics |
| 00:14:22 | How to measure the instrument response time? |
| 00:15:10 | How to measure the instrument response time? |
| 00:15:38 | Exponential fit of each return to baseline |
| 00:16:50 | Influence of the instrument response time |
| 00:18:12 | Is there kinetic information in my data? |
| 00:18:26 | Is there a variation of the equilibration time? |
| 00:19:09 | Simple consequence: compare the titration curve with the envelope of the injection curves |
| 00:21:03 | AFFINImeter |
| 00:21:43 | The simplified kinITC method: kon & koff with one click in the software AFFINImeter |
| 00:22:03 | Evolution of the equilibration time during a titration experiment |
| 00:22:28 | Evolution of the equilibration time during a titration experiment |
| 00:23:16 | Illustration with a simple case Carbonic anhydrase + inhibitor (4CBS) |
| 00:23:41 | Results for carbonic anhydrase at several temperatures |
| 00:24:33 | ETC-based kinITC |
| 00:25:08 | ETC-based kinITC |
| 00:25:30 | AFFINImeter |
| 00:25:52 | Comparison of results with SPR |
| 00:27:15 | Comparison of results with SPR |
| 00:28:40 | Take-home message |
| 00:29:14 | Contact Information |
| 00:30:34 | For further information on kinITC, please see the following resources: |
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.