Abstract

Electrochemical methods provide unique opportunities for non-traditional synthesis and analytical studies. Using electrochemistry, we are able to synthesize individually addressable molecular surfaces on microelectrode arrays and then utilize the surface to probe interactions between receptors and their biological targets. To date, cyclic voltammetry signals can be used to detect biological interactions between proteins and small molecules on the surface of an array, however those signals are often amplified by the presence of a large number of small molecules by any given electrode. This amplification makes it difficult to obtain binding data using the arrays that can be compared with existing literature data because when amplified, a signal can be shifted to a point where the binding curve cannot be observed. Therefore, it is extremely important to develop methods that allow us to calibrate the signals produced and recorded on the microelectrode arrays and optimize those signals in a reproducible way. For this reason, the research in this dissertation explores how the chemical properties of the functionalized array surface influences the binding signal. This work provides a “how-to” guide for troubleshooting analytical binding experiments in order to assess and establish the proper conditions to observe the binding activity of biological targets. We do this by adjusting the coverage of the functionalized surface, adding a linker to the ligand to ensure accessibility by the receptor, and optimizing the mediator that is used to carry the protein and produce the current measured at the electrodes. In the end, a G-protein and R6A pair are used to confirm the success of the model system we developed for monitoring real-time interactions of biological molecules.

Committee Chair

Kevin D. Moeller

Committee Members

Vladimir Birman

Degree

Doctor of Philosophy (PhD)

Author's Department

Chemistry

Author's School

Graduate School of Arts and Sciences

Document Type

Dissertation

Date of Award

Spring 5-15-2022

Language

English (en)

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