DOI

https://doi.org/10.25772/PQTA-BX09

Author ORCID Identifier

https://orcid.org/0000-0001-7356-0429

Defense Date

2020

Document Type

Dissertation

Degree Name

Doctor of Philosophy

Department

Chemical and Life Science Engineering

First Advisor

Gregory Triplett

Second Advisor

Christina Tang

Third Advisor

Hu Yang

Fourth Advisor

Mo Jiang

Fifth Advisor

Robert Tombes

Abstract

Diseases and disorders in the human body are considered abnormalities of proper cellular function. Understanding the signal transduction mechanisms that cause these abnormalities is crucial to developing earlier detection methods, better treatment options and effective cures. While current diagnostic procedures are powerful tools in diagnosing diseases, they are ineffective in informing physicians on the real-time behavior of the signal transduction mechanisms associated with diseases and disorders. Currently, disease progression is monitored over time through routine patient visits and testing by one or more of the above techniques. By developing an approach that can monitor structural and conformational changes of proteins during biochemical reactions we can provide insight towards signal transduction mechanisms. This study successfully monitored the phosphorylation of Casein through catalysis of PKA and phosphate donation from adenosine triphosphate (ATP) in a 3D printed microfluidic device that was integrated with confocal Raman spectroscopy. The development of an adaptive PCA algorithm identified variance between the POx vibrational modes at 1090 cm-1,1329 cm-1, 1411 cm-1, 1506 cm-1 and 1558 cm-1 present in ATP. The protein phosphorylation reaction was monitored in real-time by detecting the change of the POx vibrational mode at 1556 cm-1 that appears in ATP during the reaction progression. This research also enables future use on numerous other biochemical reactions utilizing Raman spectroscopy. Additionally, this work enables translation of this research to analyze biochemical reactions intracellularly.

Rights

© The Author

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

5-19-2020

Available for download on Wednesday, May 19, 2021

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