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biochemtest3-ligandbinding-SPR

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What are advantages of SPR
real-time, unique-high quality data, simple assay, small amount of sample only needed
plasmon (2)
a hybridization of conducting electrons and photons
plasmon
oscillation of the conduction electrons in a metal.
magic angle
total internal reflection
What phenomenon occurs with SPR?
electron charge density wave
What information can the sensorgram tell you?
association, concentration, kinetics, dissociation, regeneration
What are the three parts to SPR?
1. Biacore sensor chip 2. Integrated microfluid cartridges 3. Surface plasmon resonance detection
Kinetics is need for finding:
structural changes, characterization of biopharmacuetical products, assays based on affinity, purification schemes
SPR
Surface Plasmon Resonance
How is binding constant determined with SPR
bait ligand coated to gold surface of SPR crystal, prey ligand flows over bait layer and binds, SPR signal changes to new equilibrium, solution without prey is applied, new equilibrium reached. The Voff and Von can be used to calculate the binding constant.
Energy and momentum are transformed to
incident photons into surface plasmons
Quantitative measurments at the Biacore sensor chip are dependent on:
the immobilization of a target molecule to the sensor chip surface
SPR affected by
coupling of the gold layer. Binding of bait and prey affects the total internal reflection angle change
SPR can tell you
yes/no data, how much, how strong, how fast
Resonance is due to
energy and momentum

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