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Study conformational changes of prion protein by Dual polarization interferometry (DPI) (CAT#: STEM-MB-0381-WXH)

Introduction

Bovine spongiform encephalopathy, scrapie and Creutzfeldt–Jakob disease are examples of prion diseases. These diseases are sometimes referred to as transmissible spongiform encephalopathies (TSEs). Prion diseases require the expression of a normal host protein, termed the prion protein, and its conversion to an abnormal isoform. The abnormal isoform of the prion protein that accumulates in disease is termed PrPSc and is tightly associated with the infectious agent of TSEs .




Principle

Dual polarization interferometry (DPI) is an analytical technique that allows the simultaneous determination of thickness, density, and mass of a biological layer on a sensing waveguide surface in real time. DPI focuses laser light into two waveguides. One of these functions as the "sensing" waveguide having an exposed surface while the second one functions to maintain a reference beam. A two-dimensional interference pattern is formed in the far field by combining the light passing through the two waveguides. The DPI technique rotates the polarization of the laser, to alternately excite two polarization modes of the waveguides. Measurement of the interferogram for both polarizations allows both the refractive index and the thickness of the adsorbed layer to be calculated. These measurements can be used to infer conformational information about the molecular interactions taking place, as the molecule size (from the layer thickness) and the fold density (from the RI) change.

Applications

Study the mechanism by which PrPSc is generated and the differences in strains of prion disease.

Procedure

1. Setting of dual polarization interferometry
2. Preparing the DPI sensor chip
3. Immobilization of target on DPI biosensor
4. Reagent was injected to react
5. Quantitative analysis

Materials

• DPI biosensor
• DPI sensor chip