GE HEALTHCARE Biacore T100, Руководство пользователя

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9 Kinetics and affinity analysis
9.2 Evaluating kinetics and affinity in single mode
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Biacore T100 Software Handbook BR-1006-48 Edition AE
9.2.2 Multiple ligand densities
Analysis of the same analyte concentration series over multiple ligand densities
can provide more robust fitting than a single ligand density. The kinetic and
steady-state affinity fitting functions support simultaneous analysis of up to five
sets of data with independent values for R
max
, returning a single set of rate
constants for the whole combined data set. Analyses over multiple ligand
densities may be performed in separate runs that are combined with the
Append file function, but should preferably be performed on multiple flow cells
in the same run to ensure that the experimental conditions are comparable as
far as possible.
To set up evaluation of the same sample series over multiple ligand densities,
click
Multiple Rmax in the first dialog box for kinetics and affinity evaluation. A
panel for data subsets representing different R
max
values opens to the left of the
curve table.
Click Add to add a new data subset. If you have multiple sets of the currently
chosen curve type in the evaluation session (e.g. multiple reference subtracted
curves), the next curve in the list is assigned to the new data subset. If there are
no more curves of the same type available, the new set will be a copy of the
most recently created subset. You can add up to five data subsets, representing
five ligand densities. The same sample name and analysis temperature apply to
all subsets.
Click on a subset to manage the contents of the subset. You cannot mix subsets
that use different samples or different temperatures.