Chemically Specific Protein Immobilization Strategies onAnalytical Surfaces

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

Dembowy, Szymon ; Daniel L. Feldheim, Committee Member,Stefan Franzen, Committee Co-Chair,Christopher B. Gorman, Committee Chair,Dembowy, Szymon ; Daniel L. Feldheim ; Committee Member ; Stefan Franzen ; Committee Co-Chair ; Christopher B. Gorman ; Committee Chair

University:North Carolina State University

关键词: protein immobilization;    Au surface;    Ni-NTA;    PM-IRRAS;    cyclic voltammetry;    non-specific protein binding;   

Others  :  https://repository.lib.ncsu.edu/bitstream/handle/1840.16/325/etd.pdf?sequence=1&isAllowed=y

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【 摘 要 】

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.