Date of Award

Summer 8-2021

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Molecular Science and Nanotechnology

First Advisor

Gergana G. Nestorova

Abstract

Exosomes are extracellular, cell signaling microvesicles that contain unique genomic and proteomic signatures reflective of the host cell’s pathophysiological conditions. In recent years, the study of exosomes has increased tremendously because many have been recognized as molecular biomarkers with the potential to advance methods of disease diagnostics and therapeutics as well as contribute to physiological analyses of multiple organism types. With the promising potential that exosomes offer to the field of molecular biology, it is vital to establish an efficient and consistently reliable mechanism of exosome isolation from biological samples. Many isolation techniques currently available typically yield exosome samples with numerous contaminants, making them low in purity for exosome content. Our lab has developed a novel technology for solid-phase exosome purification directly from biological samples. This study utilizes SDS-PAGE and fluorescent imaging analysis to assess the specificity of the ExoSense microprobe-based exosome isolation technology. The proteomic profile generated from the SDS-PAGE shows fewer bands with a cleaner background for the microprobe-based sample compared to the traditional polymer precipitated exosome proteins, suggesting an exosome population higher in purity from the microprobes than what the polymer reagent provides. Fluorescent imaging resulted in distinct differences between the control and exosome-specific groups, indicating an exosome-specific population captured using the probes. Results from this study can be used collectively to validate the specificity of the novel ExoSense exosome capture technology. Future studies can be applied for optimizing the technology for commercial applications.

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