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Optical Spec 7 - Surface Plasmon Resonance

Optical Spec 7 - Surface Plasmon Resonance

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Published by Miles Nsg

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Published by: Miles Nsg on Jan 11, 2011
Copyright:Attribution Non-commercial

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12/25/2012

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Physical Biochemistry Surface Plasmon Resonance[Page 1]  A technique to study binding of analytes to ligandsThe analyte flows through the flow cell (F)Immobilised ligand coats substrate (S) which is attached to a prism (P)The light source (L) illuminates the substrate with a thin wedge of light TIR occurs at the surface of the substrate A detector array (D) records the reflected wedge of light The detector is made up of small cells arranged along a lineEach detector will detect light being reflected at a certain angle At most angles there would be constant reflection, but at certainangles the amount of reflected light would fall, due to surface plasmon resonance.When a ligand is bound to the substrate, the SPR dip will occur at a certain angle, when analyte moleculesbind to the ligand, the dip will shift. We measure how this µdip¶ shifts with angle as a function of time.The Sensor Chip: A 3D matrix has the ligand immobilised onto it. This increases the density of the ligand / surface area. Applications:The change in SPR is directly proportional to mass being immobilised Useful for equilibrium and kinetic measurementsNo labelling required but has to be immobilised  Advantages:
 
No labelling
 
Real time (fast measurements, sub-second time scale)

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