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Permanent link (DOI): https://doi.org/10.7939/R38S4S

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Development of Valve-based Microchip for Proteomics Open Access

Descriptions

Other title
Subject/Keyword
fractionation
valve-based microchip
bead packing
Proteomics
electrospray ionization mass spectrometry
integration
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Lu, Qingye
Supervisor and department
Harrison, D. Jed (Chemistry)
Examining committee member and department
Wheeler, Aaron (Chemistry, University of Toronto
Klobukowski, Mariusz (Chemistry)
Li, Liang (Chemistry)
McDermott, Mark T. (Chemistry)
Weiner, Joel (Biochemistry)
Department
Department of Chemistry
Specialization

Date accepted
2010-01-28T21:23:39Z
Graduation date
2010-06
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
Interest in microfluidic platforms has surged as an alternative for sample preparation in the past two decades, with the potential for miniaturization, portability, automation, integration and parallelism driving this research. However, it is still very challenging to develop an integrated microfluidic device for proteomic preparation for mass spectrometry analysis. My thesis work is focused on the development of a valve-based microfluidic platform interfaced with electrospray ionization mass spectrometry for multiplexed proteomic analysis. First, techniques are developed for the fabrication and packing of multiple beds in a polydimethylsiloxane (PDMS) microdevice, which is compatible with the integration of multilayer microvalves. A soft lithography technique was used to fabricate stable weirs in microchips and new bead introduction techniques were explored for the elimination of bead introduction channels in the design. Such a combination provides a convenient, efficient and effective way for multiple bed preparation in a complex design. Next, detailed studies were carried out on the design parameters and performance of multilayer PDMS microvalves in the presence of high electric fields. These studies guided the integration of electrophoresis methods with valve-based fractionation. Finally, a coupled CE-fractionation-SPE-ESIMS peptide analysis on a totally integrated valve-based microchip is presented. We show the design and operation of a system that performs electrokinetic separation, followed by fractionation into multiple channels, SPE extraction and sample cleanup on packed reaction beds, using a multiplexed, hydraulically valved system, with subsequent mass spectral (MS) analysis. This coupled multiple channel CE-Fractionation-SPE-ESIMS platform on valve-based microchip was successfully applied to peptide analysis.
Language
English
DOI
doi:10.7939/R38S4S
Rights
License granted by qingye lu (qingye@ualberta.ca) on 2010-01-28T03:40:16Z (GMT): Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of the above terms. The author reserves all other publication and other rights in association with the copyright in the thesis, and except as herein provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
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