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Transcript
Examensarbete Activity of carbonic anhydrase II in presence of nanoparticles. Coating effects The interaction of condensed materials with biological systems has long been a subject of scientific and practical interest. The increase development and exposure of drug delivery systems requires a thoughtful approximation to the possible consequences. Even though those aggregates have a specific function, drug delivery, biocatalysis, etc.; unexpected effects can rise from the exposure of those systems to living organism and in particular to the performance of highly functional molecules as enzymes. An important feature of nanomaterials is the protective coating. Many nanoparticles need an extra organic layer in order to gain colloidal stability in solution and avoid coalescence and precipitation. Therefore, for the same functional core material a series of coatings can be found depending on the application. The primary interaction of surface active molecules such as enzymes will consequently be the stabilizing layer. As a consequence of this, the kinetic effect upon adsorption must be related to the physicochemical characteristics of this coating. The first step in this project is to analyze the effect of the interaction between colloidal structures such as metallic nanoparticles and enzymes. The activity and performance of enzymes can be extremely sensitive to small changes in protein conformation. The association to surfaces can cause such modifications and severely modify the protein functioning. Using time-resolved spectroscopic methods you will investigate how kinetics effects correlate with changes in conformation upon binding. HCA II Au@PSS NPs The association process between carbonic anhydrase II and gold polyelectrolyte coated nanoparticles will be studied as well as the changes in enzyme conformation. These changes will be correlated to the kinetics effects observed. The same enzymatic reaction will be analyzed in a system mimicking a biological fluid. A neutral (non- reacting) protein would be introduced in order to study the effect of the competition between the proteins in solution for the particle surface. The final goal is to relate the on and off rates and the binding constants with the overall kinetic picture. Techniques. Ø Ø Ø Ø Ø UV-spectroscopy CD spectroscopy Dynamic light scattering Isothermal calorimetry Surface Plasmon resonance For further information, please contact me at [email protected]