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9 Kinetics and affinity analysis
9.1 Requirements for kinetics and affinity evaluation
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Biacore T100 Software Handbook BR-1006-48 Edition AE
Figure 9-1. In multi-cycle kinetics and affinity determinations, each sample is injected in
a separate cycle. The concentration series is presented as an overlay plot aligned at the
start of the injection in the evaluation software.
In single-cycle determinations, the samples are injected sequentially in the same cycle.
Arrows in the illustrations mark the start of sample injections.
9.1 Requirements for kinetics and affinity evaluation
The minimum requirements for evaluation of kinetics or affinity are one cycle
with a
Sample injection in an assay step with purpose Sample , and with the
sample concentration in the keyword Conc . If the concentration is not given in
molar-based units, the keyword MW must also be included with a value for the
molecular weight. For method-based runs, the method must be correctly
constructed as described in Section 5.10.2: if necessary, the keyword table can
be edited so that the conditions are met in full (see Section 6.6). Note however
that the command type cannot be edited in the keyword table. Refer to
Chapter 5 for details of how to construct methods in Method Builder.
The recommended minimum conditions for kinetic and affinity evaluation are:
• a concentration series of analyte with at least four non-zero
concentrations
• at least one blank cycle consisting of zero concentration sample (for
single-cycle kinetics the blank cycle must replicate the sequence of
injections in the analysis cycle)
• for multi-cycle kinetics, duplicate determinations for at least one non-zero
concentration.
These conditions are recommended but not mandatory in the Kinetics/Affinity
wizard.