Identifier

etd-11252009-165756

Degree

Doctor of Philosophy (PhD)

Department

Chemistry

Document Type

Dissertation

Abstract

The need for rapid, portable and high-throughput systems in proteomics is now prevalent because of demands for generating new protein-based disease biomarkers. However, 2-D protein profile patterns are lending themselves as potential diagnostic tools for biomarker discovery. It is difficult to identify protein biomarkers which are low abundant in the presence of highly abundant proteins, especially in complex biological samples like serum. Protein profiles from 2-D separation of the protein content of cells or body fluids, which are unique to certain physiological or pathological states, are currently available on internet databases. In this work, we demonstrate the ability to separate a complex biological sample using low cost, disposable, polymer-based microchips suitable for a multidimensional techniques that employed sodium dodecyl sulfate micro-capillary gel electrophoresis (SDS µ-CGE) in the 1st dimension and micellar electrokinetic capillary chromatography (MEKC) or microemulsion electrokinetic capillary chromatography (MEEKC) in the 2nd dimension. The peak capacity generated by this microchip technique was about 3-fold greater compared to conventional 2-D separation methods and the complete separation time was 60X faster. To minimize electroosmotic flow effects, we dynamically coated the channels with methylhydroxyethyl cellulose. Proteins were detected by laser-induced fluorescence following their labeling with dyes. To mitigate challenges posed by labeling the proteins, we investigated the use of a label-free technique that relied upon conductivity measurements. Preliminary data are presented on the fabrication of on-chip electrodes using a conductive SU-8 polymer via lithography.

Date

2009

Document Availability at the Time of Submission

Secure the entire work for patent and/or proprietary purposes for a period of one year. Student has submitted appropriate documentation which states: During this period the copyright owner also agrees not to exercise her/his ownership rights, including public use in works, without prior authorization from LSU. At the end of the one year period, either we or LSU may request an automatic extension for one additional year. At the end of the one year secure period (or its extension, if such is requested), the work will be released for access worldwide.

Committee Chair

Soper, Steven

DOI

10.31390/gradschool_dissertations.1816

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