The research presented in this work explores the application of microfluidics to the field of proteomics through the design of a multi-channel microfluidic platform and the investigation of individual components of the system. The design of this microfluidic device allows the integration of several protein sample preparation steps for automated electrospray ionization mass spectrometric (ESI-MS) analysis, including protein separation, fractionation and collection, preconcentration and cleanup, and protein digestion.
In order for the multi-channel system to function properly, I first evaluated each individual component of the device. Several areas were explored: (i) optimization of polymer monolith for solid-phase extraction (SPE) preconcentration; (ii) investigation of cationic coatings for microchannel surface modification to facilitate positive electrospray of peptides and proteins for chip-MS coupling; (iii) combination of the hydrophobic monolith and the PolyE-323 coating in a single channel device for on-chip SPE and on-bed tryptic digestion with on-line coupling to ESI-MS.
Multiplexed microfluidic devices for protein analysis, which integrate a series of microfluidic features, were then designed, built and tested. The multiplexed microfluidic architecture employed a separation channel, a fractionator, an array of microchambers to accommodate monolithic polymer for SPE preconcentration, and an elution channel for the detection of eluted sample using fluorescence detector or mass spectrometer. The performance of the multiplexed devices for integration of multiple analytical steps was explored with sequential fractionation, collection, and elution of fluorescent sample, evaluating the ability to trap and release individual fractions without cross-contamination.
Thorough analysis of each of the individual components on the multiplexed microfluidic platform provides valuable insights into the design of such systems, which brings us closer to our final goal of a proteomic processing microchip.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:AEU.10048/925 |
| Date | 06 1900 |
| Creators | Hua, Yujuan |
| Contributors | D. Jed Harrison (Chemistry), Charles A. Lucy (Chemistry), Liang Li (Chemistry), Jonathan G. C. Veinot (Chemistry), Christine M Szymanski (Biological Sciences), David Sinton (Mechanical Engineering, University of Victoria) |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Format | 8043544 bytes, application/pdf |
| Relation | Yujuan Hua, D. Jed Harrison* Proceedings of MicroTAS 2009, The 13th International Conference on Miniaturized Systems for Chemistry and Life Sciences, Jeju Island, Korean, p. 1719-1721., Yujuan Hua, Zhen Wang, D. Jed Harrison* Proceedings of MicroTAS 2007, The 11th International Conference on Miniaturized Systems for Chemistry and Life Sciences, Paris, France, p. 1234-1236. |
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