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Study of lysozyme on adsorption solid surfaces by Dual polarization interferometry (DPI) (CAT#: STEM-MB-0431-WXH)

Introduction

Lysozyme is an enzyme that is able to lyse bacterial cell membranes and thus serve as an antimicrobial agent in foods. Lysozyme occurs naturally in egg white, but the purified enzyme is used on occasion as an additive in other foods, especially soft cheeses. Lysozyme is a well-documented egg white allergen. Lysozyme can be utilized as model protein in crystallization experiments and X-ray structure analysis.
Proteins accumulate at interfaces, a property that can be both a practical asset and a problem. New biotechnological methods of protein production depend on protein interracial properties in downstream protein purification and separation. Furthermore, the adsorption of proteins at solid/liquid interfaces has enabled the development of diverse biomedical applications, such as biosensors, immunological tests, and drug-delivery schemes. In the biomaterial field, protein adsorption is much less desirable because it can elicit adverse host responses such as blood coagulation and complement activation.




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 of lysozyme on adsorption solid surfaces.
Biosensors.
Biomaterial.
Immunological tests.
Drug-delivery schemes.

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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