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