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Design and fabrication of a continuous flow mixer for investigating protein folding kinetics using focal plane array Fourier transform infrared spectroscopy

This thesis presents the design, fabrication, and testing of a micromixing device intended for use in investigating protein dynamics on a microsecond timescale by Fourier transform infrared (FTIR) spectroscopy. Numerical modeling of flow was implemented to predict the influence of flow rates and geometric variations on mixing performance in three passive mixers. The simulation models were validated by experimental measurements using optical and infrared detection. The optimum level of mixing was observed in a multi-lamination mixer that combined thin filaments of differing fluids in an alternating manner. The multi-laminates were transferred onto polished calcium fluoride infrared-transparent optical windows by lithographic processing of an Epon-based polymer, SU-8. A rigid seal between two microchannels was accomplished through thermal bonding of an unexposed resist layer, which acted as a thermal epoxy under the influence of temperature. The multi-lamination mixer was used to study the changes in the secondary structure of beta-Lactoglobulin in deuterated phosphate buffer under varying physicochemical conditions by time-resolved FTIR spectroscopy using focal plane array detection. Upon a pH jump from pH 2 to neutral pH, a gradual loss of alpha-helical content, accompanied by an increase in random coils and turns was observed within 2 ms of mixing. In a second kinetic experiment, mixing of a neutral-pH solution of beta-Lactoglobulin with a 60% trifluoroethanol solution resulted in the formation of an alpha-helical intermediate with an accompanying increase in intramolecular beta-sheet structure within 500 mus of mixing. These results indicate that the multi-lamination mixer designed and fabricated in this study is well suited for investigations of protein dynamics on the micro- to millisecond timescale by time-resolved FTIR spectroscopy.

Identiferoai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.116093
Date January 2008
CreatorsHaq, Moeed.
PublisherMcGill University
Source SetsLibrary and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada
LanguageEnglish
Detected LanguageEnglish
TypeElectronic Thesis or Dissertation
Formatapplication/pdf
CoverageMaster of Science (Department of Food Science and Agricultural Chemistry.)
RightsAll items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated.
Relationalephsysno: 003164293, proquestno: AAIMR67035, Theses scanned by UMI/ProQuest.

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