GE HEALTHCARE Biacore T100, Handbook

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9 Kinetics and affinity analysis
9.4 Quality assessment for kinetics evaluation
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Biacore T100 Software Handbook BR-1006-48 Edition AE
9.4.3 Components of the fit
Choose Tools:Components from the fitted results window to display a plot
showing the contribution of components in the interaction model to the fitted
curve. Choose which cycle to display in the selector bar.
The example illustrated here is taken from a fitting to a bivalent analyte model
(see Section 9.8.2), and shows clearly how the component AB2 (analyte attached
to the surface through both binding sites) is displaced by AB as the interaction
progresses.
9.4.4 Check kinetic data
Kinetic constants obtained from the fitting procedure are only significant if the
observed binding is not seriously limited by mass transport of analyte to the
surface (see Section 9.8). For 1:1 fitting results, you can check whether mass
transport is limiting or not using the
Check Kinetic Data function.
Choose Tools:Check Kinetic Data to open a dialog that displays simulated
sensorgrams based on the fitting results, with the interaction rate constants k
a
and k
d
varied in parallel (so that the affinity constant, remains unchanged). If
curves do not shift as values for k
a
and k
d
are changed, this means that the
actual values are not important for the fitting, and the curves do not contain
kinetic information. Conversely, if the simulated curve shape changes as the
values of k
a
and k
d
are varied, the fitting is dependent on the actual values and
the curves do contain kinetic information.
This tool is only available for results obtained with the 1:1 fitting model.