New dimensions in proteomic diagnostics using Raman spectroscopy
Abstract
High-quality normal Raman spectra are derived from protein solutions with concentrations down to 1 μM and 1 fmol of protein nondestructively probed within the excitation laser beam. These results are obtained using a drop coating deposition Raman (DCDR) method in which the solution of interest is microdeposited (or microprinted) on a compatible substrate, followed by solvent evaporation and backscattering detection. Representative applications include the DCDR detection of insulin derived from an HPLC fraction, nondestructive DCDR followed by MALD1-TOF of lysozyme, the DCDR detection of protein spots deposited using an ink-jet microprinter, the identification of spectral differences between glycan isomers of equal mass (such as those derived from post-translationally modified proteins), DCDR detection of spectral differences in insulin variants, DCDR detection of phosphorylation sites in peptides and/or proteins, detection of insulin conformational changes involved in fibrillation, and protein-ligand binding DCDR studies.
Degree
Ph.D.
Advisors
Ben-Amotz, Purdue University.
Subject Area
Analytical chemistry
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