Date of Award

1-1-2019

Language

English

Document Type

Master's Thesis

Degree Name

Master of Science (MS)

College/School/Department

Department of Chemistry

Content Description

1 online resource (ix, 31 pages) : illustrations (some color)

Dissertation/Thesis Chair

Igor Lednev

Committee Members

Mehmet Yigit

Keywords

Disease diagnostics, RNA Binding Proteins, SERS, Single molecule, Raman effect, Surface enhanced, Raman spectroscopy, Proteins, Spectrum Analysis, Raman, protein

Subject Categories

Biochemistry | Chemistry

Abstract

A two-step process of protein detection at a single molecule level using Surface Enhanced Raman Spectroscopy (SERS) was developed as a new platform for medical diagnostics in this proof-of-concept study. First, a protein molecule was bound to a linker in the bulk solution and then this adduct was chemically reacted with the SERS substrate. Traut’s Reagent (TR) was used to thiolate Bovine serum albumin (BSA) in solution followed by chemical cross linking to a gold surface through a sulfhydryl group. A Glycine-TR adduct was used as a control sample to identify the protein contribution to the SER spectra. Gold SERS substrates were manufactured by electrochemical deposition. Solutions at an ultralow concentration were used for attaching the TR adducts to the SERS substrate. Samples showed a typical behavior of a single molecule SERS including spectral fluctuations, blinking and Raman signal being generated from only selected points on the substrate. The fluctuating SER spectra were examined using Principle Component Analysis. This unsupervised statistics allowed for selecting spectral contribution from protein moiety indicating that the method was capable of detecting a single protein molecule. Overall, it was demonstrated, that the developed two-step methodology has the potential as a new platform for medical diagnostics.

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