2024-03-28T17:15:21Zhttps://keep.lib.asu.edu/oai/requestoai:keep.lib.asu.edu:node-1502572021-08-30T18:50:26Zoai_pmh:all150257
https://hdl.handle.net/2286/R.I.14280
http://rightsstatements.org/vocab/InC/1.0/
All Rights Reserved
2011
xiv, 152 p. : ill. (some col.)
Doctoral Dissertation
Academic theses
Text
eng
Weiss, Noah Graham
Hayes, Mark A.
Garcia, Antonio
Ros, Alexandra
Arizona State University
Partial requirement for: Ph.D., Arizona State University, 2011
Includes bibliographical references
Field of study: Chemistry
Bioanalytes such as protein, cells, and viruses provide vital information but are inherently challenging to measure with selective and sensitive detection. Gradient separation technologies can provide solutions to these challenges by enabling the selective isolation and pre-concentration of bioanalytes for improved detection and monitoring. Some fundamental aspects of two of these techniques, isoelectric focusing and dielectrophoresis, are examined and novel developments are presented. A reproducible and automatable method for coupling capillary isoelectric focusing (cIEF) and matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) based on syringe pump mobilization is found. Results show high resolution is maintained during mobilization and &beta-lactoglobulin; protein isoforms differing by two amino acids are resolved. Subsequently, the instrumental advantages of this approach are utilized to clarify the microheterogeneity of serum amyloid P component. Comprehensive, quantitative results support a relatively uniform glycoprotein model, contrary to inconsistent and equivocal observations in several gel isoelectric focusing studies. Fundamental studies of MALDI-MS on novel superhydrophobic substrates yield unique insights towards an optimal interface between cIEF and MALDI-MS. Finally, the fundamentals of isoelectric focusing in an open drop are explored. Findings suggest this could be a robust sample preparation technique for droplet-based microfluidic systems. Fundamental advancements in dielectrophoresis are also presented. Microfluidic channels for dielectrophoretic mobility characterization are designed which enable particle standardization, new insights to be deduced, and future devices to be intelligently designed. Dielectrophoretic mobilities are obtained for 1 µm polystyrene particles and red blood cells under select conditions. Employing velocimetry techniques allows models of particle motion to be improved which in turn improves the experimental methodology. Together this work contributes a quantitative framework which improves dielectrophoretic particle separation and analysis.
Chemistry
Dielectrophoresis
Gradient separation
Isoelectric focusing
New developments in isoelectric focusing and dielectrophoresis for bioanalysis