GE HEALTHCARE Biacore T100, Handbook

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Biacore T100 Software Handbook BR-1006-48 Edition AE
71
Application wizards 4
changed does not interfere with the first measurements. Start-up cycles will be
run at each temperature.
Check Run solvent correction to include solvent correction cycles in the run.
Solvent correction adjusts response values for the effects of varying bulk
refractive index contribution between samples, and may improve the quality of
the results for analyses using small organic analytes that require dimethyl
sulfoxide (DMSO) in the sample buffer to maintain solubility. The principles and
application of solvent correction are described in Section 6.7.
Set the required number of injections per cycle and the frequency of solvent
correction cycles. The default settings use 8 injections per cycle and repeat the
solvent correction cycle every 15 sample cycles. Separate solvent correction
cycles will be run at each temperature.
Enter the temperatures at which the measurements are to be performed. For
most purposes, 5-7 temperatures will be adequate: fewer points make the
determination of thermodynamic parameters uncertain, while more points
increase the run time. Distribute the points evenly over the widest temperature
interval that the ligand and analyte tolerate. Start from the lowest temperature
to minimize the time needed for temperature equilibration between
measurements (increasing the analysis temperature takes less time than
decreasing it). The system will wait for a stable temperature between each
determination. As an additional control, you may want to include a replicate of
the first temperature at the end of the run.
Note: For robust determination of standard thermodynamic parameters (
Δ
G°,
Δ
H° and
Δ
S°), arrange the analysis temperatures so that 25°C is in the
middle of the range (e.g. 10–40°). See Section 10.1 for further details.
Specify the sample compartment temperature, or check Vary with analysis
temperature to change the sample compartment temperature automatically
when the analysis temperature is changed.
Step 3. Injection parameters
Specify the injection parameters for each injection in the cycle (see
Section 4.2.3). The sample injection for thermodynamics measurement has an
additional setting for dissociation time. This is the time for which dissociation will
be monitored after the end of the injection without disturbances from flow
system washing procedures.
Step 4. Samples
Enter the details of the samples. This dialog box is equivalent to the
concentration series dialog for kinetics determination (see Section 4.5.3, step 4).
Separate rack positions will be created for each sample at each temperature.