Surface Plasmon Resonance SPR

Question 1

Experimental methods to determine protein ligand interactions

Answer 1

Fluorescence Polarization
Surface Plasmon Resonance
Isothermal Titration Calorimetry
NMR
MS

Question 2

Steps SPR

Answer 2

1) Buffer flow (baseline)
2) Ligand flow. Rate of association monitored
3) Equilibirum bound/unbound ligand
4) Buffer flow to dissociate
5) Restore baseline

Question 3

How SPR

Answer 3

Protein immobilized onto metal surface/chip
Ligand flows through chip
Light source
Detector
Binding event measure as
- change mass
- change refractive index
- changebin reflaction angle

Question 4

What do you get SPR

Answer 4

Kd
Rates of association dissociation