Author ORCID Identifier
Semester
Spring
Date of Graduation
2026
Document Type
Dissertation
Degree Type
PhD
College
Eberly College of Arts and Sciences
Department
Chemistry
Committee Chair
Lisa Holland
Committee Member
Stephen Valentine
Committee Member
Peng Li
Committee Member
Harry Finklea
Committee Member
Mariette Barbier
Abstract
Capillary electrophoresis as a tool for biomolecule analyses provides fast, high efficiency separations consuming only nanoliters of sample. Coupled to mass spectrometry, insight of analyte structure is gained which is not always easily determined by capillary electrophoresis alone. The work presented in the dissertation applies capillary electrophoresis – mass spectrometry coupled by a novel vibration based ionization source for the analysis of proteins, glycoproteins, and amyloids.
Protein detection with capillary electrophoresis is not straight forward. Positively charged analytes adsorb to the capillary and resolution between similarly charged large molecules can prove difficult. Capillary coatings were explored to both neutralize and change the charge of the capillary wall to increase the efficiency of the protein separations and expand the typical analyte set available when using a coated capillary. Vibrating sharp-edge spray ionization was also demonstrated the ionization of intact protein complexes.
Heterogeneous protein glycosylation can be incredibly difficult to characterize due to variation in glycan structure such as branching, sialylation, fucosylation, and the possibility of isomeric structures. Typically, glycans are cleaved from the protein and analyzed as a general population, however, that information is near impossible to piece back into a glycosylation profile of an intact glycoform. Here, nanoliter volume reactions were accomplished in-line during the separation to interact with or modify glycan structure for the elucidation the overall glycosylation of all detected glycoforms.
Since the outbreak of SARS-CoV-2, it has been reported that infection has resulted in the onset or acceleration of neurodegenerative disease. Amyloidogenic peptides of the SARS-CoV-2 Spike protein have been identified and reported to influence the aggregation of amyloid-β(1-42), a cause of Alzheimer’s diseases, as well as other aggregation prone proteins linked to neurodegeneration. Capillary electrophoresis – vibrating sharp-edge spray ionization – mass spectrometry was used to detect heteromeric aggregation between amyloid-β(1-42) and synthetic aglycosylated Spike protein amyloids.
Recommended Citation
Witzel, Makenzie, "Capillary Electrophoresis – Vibrating Sharp-edge Spray Ionization – Mass Spectrometry for Protein Analysis, Glycoprotein Characterization, and Detection of Amyloid Heteromeric Aggregation" (2026). Graduate Theses, Dissertations, and Problem Reports. 13234.
https://researchrepository.wvu.edu/etd/13234