GE HEALTHCARE Biacore T100, Handbook

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Biacore T100 Software Handbook BR-1006-48 Edition AE
73
Application wizards 4
4.6.1 Mass transfer control
Each sample entered in the Samples step will be analyzed three times, at fixed
flow rates of 5, 15 and 75 µl/min, with a contact time of 1 minute and
dissociation time of 2 minutes. Use an analyte concentration that gives readily
measurable initial binding rates. Mass transfer limitation is not affected by
analyte concentration, but dependence of binding rate on flow rate may be
difficult to detect if the binding rate is too low or too high.
Note: All predefined kinetic evaluation models include a term for mass transfer,
and the original purpose of the mass transfer control experiment has
largely been superseded by functions in the evaluation software. The
control experiment may however still be useful to confirm suspected
mass transfer limitations if desired.
4.6.2 Linked reactions control
Each sample entered in the Samples step will be analyzed at a flow rate of
10 µl/min with fixed contact times of 0.5, 3 and 10 minutes and a dissociation
time of 10 minutes. Use one or more fairly high analyte concentrations,
preferably so that steady state is approached or reached within the shortest
contact time. The experiment is easiest to interpret if the interaction is
maintained at steady state for varying lengths of time, so that the starting
response for dissociation is constant.
Note: The Injection Parameters step allows you to enter a stabilization time
after each sample injection, but all other settings in this dialog are fixed in
the wizard.