The heat released or absorbed during titration of the ligand into the target solution is directly proportional to the ligand-target binding which provides a thermodynamic profile of the molecular interaction.
The integrated area of each injection peak is plotted against the molar ratio of ligand to target. The isotherm is fitted to a binding model in order to derive binding affinity (KD) and reaction stoichiometry (n).
Example of evaluation of BRD4 bromodomain 1 (BRD4-1) binding to JQ1
Upper image: Overlay of titration spectrograms of BRD4-1 protein vs. JQ1 (black) and buffer (blue).
Lower image: integrated data for BRD4-1/JQ1 binding was analyzed using one-site binding model. The best fit parameters are shown in the box. JQ1 shows binding to BRD4-1 with an estimated KD of 36 nM.
Instruments: The low volume nano ITC (TA instruments) is highly sensitive and uses less sample than the standard volume ITC instrument. The cell is constructed of inert gold with high thermal conductivity. Cylindrical cell geometry maximizes stirring efficiency and eliminates dead zones and bubbles.
Turnaround time: The study is typically completed within 2 weeks after receipt of materials. Projects are performed on a first-come-first-serve basis.
Sample requirements: Protein purity of 90% or more is required. The protein amount is project-dependent. Typically, 1-5 mg of pure protein or more will be needed.
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Screening location: The assay is performed in Malvern, PA, USA.