New Developments in Isoelectric Focusing and Dielectrophoresis for Bioanalysis

abstract: 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 o...

Full description

Bibliographic Details
Other Authors: Weiss, Noah (Author)
Format: Doctoral Thesis
Language:English
Published: 2011
Subjects:
Online Access:http://hdl.handle.net/2286/R.I.14280
id ndltd-asu.edu-item-14280
record_format oai_dc
spelling ndltd-asu.edu-item-142802018-06-22T03:02:14Z New Developments in Isoelectric Focusing and Dielectrophoresis for Bioanalysis abstract: 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. Dissertation/Thesis Weiss, Noah (Author) Hayes, Mark A (Advisor) Garcia, Antonio (Committee member) Ros, Alexandra (Committee member) Arizona State University (Publisher) Chemistry Dielectrophoresis Gradient separation Isoelectric focusing eng 167 pages Ph.D. Chemistry 2011 Doctoral Dissertation http://hdl.handle.net/2286/R.I.14280 http://rightsstatements.org/vocab/InC/1.0/ All Rights Reserved 2011
collection NDLTD
language English
format Doctoral Thesis
sources NDLTD
topic Chemistry
Dielectrophoresis
Gradient separation
Isoelectric focusing
spellingShingle Chemistry
Dielectrophoresis
Gradient separation
Isoelectric focusing
New Developments in Isoelectric Focusing and Dielectrophoresis for Bioanalysis
description abstract: 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. === Dissertation/Thesis === Ph.D. Chemistry 2011
author2 Weiss, Noah (Author)
author_facet Weiss, Noah (Author)
title New Developments in Isoelectric Focusing and Dielectrophoresis for Bioanalysis
title_short New Developments in Isoelectric Focusing and Dielectrophoresis for Bioanalysis
title_full New Developments in Isoelectric Focusing and Dielectrophoresis for Bioanalysis
title_fullStr New Developments in Isoelectric Focusing and Dielectrophoresis for Bioanalysis
title_full_unstemmed New Developments in Isoelectric Focusing and Dielectrophoresis for Bioanalysis
title_sort new developments in isoelectric focusing and dielectrophoresis for bioanalysis
publishDate 2011
url http://hdl.handle.net/2286/R.I.14280
_version_ 1718699374344667136