Chemically specific protein immobilization strategies on analytical surfaces

by 1976- Dembowy, Szymon

DEMBOWY SZYMON. Chemically Specific Protein Immobilization Strategies on Analytical Surfaces. (Under the direction of Professor Christopher B. Gorman and Professor Stefan Franzen) Chemically specific protein immobilization strategies on surfaces were explored. The goal of this work is to capture and bind proteins to the surface without losing their physiological properties. This approach permits to express functions of proteins after immobilization process. Using nitrilotriacetic acid (NTA) linkers and polyhistidine-tagged proteins, several synthetic strategies for presenting these linkers to the surface are illustrated. Gold was used as a primary surface. Data was also obtained for other potentially useful surfaces, including Indium-Tin Oxide (ITO) and germanium. Variable Angle Reflectance FTIR, Polarization- Modulation-IR Reflection Absorption Spectroscopy and Atomic Force Microscopy were used to characterize assembly process and structures on the surfaces. In-situ system assembly optimization was attempted by using a Ferrocene-based electroactive probe. Extensive study of specific and non-specific binding was conducted for a wide range of proteins.