Chemically Specific Protein Immobilization Strategies on Analytical Surfaces

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.

Author dc:creator

• Dembowy, Szymon

Advisors dc:contributor.advisor

• Daniel L. Feldheim, Committee Member
• Stefan Franzen, Committee Co-Chair
• Christopher B. Gorman, Committee Chair

• protein immobilization
• Au surface
• Ni-NTA
• PM-IRRAS
• cyclic voltammetry
• non-specific protein binding

Statement dc:rights

• I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to NC State University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report.

Dc Identifier Other

etd-10252004-145733

OAI record oai:identifier

https://repository.lib.ncsu.edu/server/oai/request?verb=GetRecord&metadataPrefix=dim&identifier=oai%3Arepository.lib.ncsu.edu%3A1840.16%2F325