ProteOn XPR36, User Manual

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ProteOn XPR36 System | C Surface Plasmon Resonance
214
• Binding kinetics — rate at which two molecules associate and
dissociate, on/off rates, association constant (k
a
), dissociation
constant (k
d
)
• Binding affinity — strength of attraction between two molecules,
equilibrium analysis (K
D
)
• Concentration determination — analyte quantitation (comparison
with standard curve)
These data elucidate many different facets of protein interactions in a variety
of biomolecular applications. These applications include the characterization
and development of antibodies, investigation of drug-target interactions,
analysis of protein interfaces, and exploration of protein complexes and
interaction cascades.
Interaction Analysis
For interaction analysis with the ProteOn XPR36 system, one interactant (the
ligand) is immobilized on the sensor chip surface, and the other interactant
(the analyte) is passed in solution over that surface. The analysis consists of a
series of steps and the requirement for each of these steps depends on
sensor chip chemistry. ProteOn Manager™ software arranges these steps into
two major groups: ligand immobilization steps and analyte interaction steps.
Ligand Immobilization Steps
• Activation (GLC and GLM sensor chips only)
• Immobilization
• Deactivation (GLC, GLH, and GLM sensor chips only)
• Stabilization (optional)
For ligand immobilization, the multichannel module (MCM) is pressed against
the sensor chip surface, forming a set of six parallel channels through which
samples and reagents are injected. The sensor chip surface is activated
(prepared for coupling) and the ligand is immobilized (bound). The remaining
(unbound) activated groups on the sensor chip surface are deactivated, and
the sensor chip surface is washed to remove nonspecifically bound molecules
(stabilization).
Analyte Interaction Steps
• Analyte injection
• Regeneration (optional)