Monitoring the interaction of lysozyme with an acrylic-based hydrogel by Dual polarization interferometry (DPI) (CAT#: STEM-MB-0458-WXH)

Introduction

The interactions of proteins with hydrophilic materials is of importance in the contact lens industry due to their implication in allergic and inflammatory reactions as well as microbial contamination which may lead to infection. Hydrogel contact lenses with high water content tend to have an open matrix structure that may influence the uptake of proteins. The molecular sizes of proteins are also thought to play a role in protein adsorption and absorption. Lysozyme has been implicated as one of the major contributors to inflammation.
Ionic monomers such as methacrylic acid (MAA) are commonly used in hydrogel contact lens materials to increase the water content of the lenses and thus increase the amount of oxygen that can reach the cornea. Previous reports have shown that protein is extensively taken up by hydrogels containing MAA.

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Principle Applications Procedure Materials Notes Related Products

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

Monitoring the interaction of lysozyme with an acrylic-based hydrogel.
Contact lens industry.

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

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