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Title page for ETD etd-09072012-084324

Type of Document Dissertation
Author Enders, Jeffrey Richard
URN etd-09072012-084324
Title Structural mass spectrometry strategies for systems biology
Degree PhD
Department Chemistry
Advisory Committee
Advisor Name Title
John A. McLean Committee Chair
Brian O. Bachmann Committee Member
David E. Cliffel Committee Member
John P. Wikswo Committee Member
  • MS
  • network dynamics
  • cellular dynamics
  • systems biology
  • mass spectrometry
  • ion mobility
  • microfluidics
  • solid phase extraction
  • IM
  • SPE
  • online SPE
Date of Defense 2012-08-30
Availability unrestricted
Systems biology seeks to describe the function of a biological system using a holistic, multiscale approach. This approach encompasses the analyses of molecular classes such as the genome, transcriptome, proteome, and metabolome (among others) of a biological system. Systems biology asserts that while understanding genes and proteins is important, the most important aspects going forward are more closely related to the system’s network structure and network dynamics. There is a substantial need for improved techniques for coupling the system on which the analysis is being performed (i.e., the organism) with the detection instrumentation (i.e., the ion mobility-mass spectrometer). Advances in systems biology measurements which focus on improved dynamic detection strategies are presented in this work. Our preliminary research shows the potential for this system to provide temporally resolved systems biology data directly from complex cellular culture populations. Precise and expedient control of the microenvironment of the MTNP with regards to input concentrations (i.e., stimuli) has been demonstrated and shows that biomolecular temporal events on the order of 5 minutes are observable with this platform. This accomplishment is critical for future stimulus/response experiments. The broad and temporally accurate detection capabilities of IM-MS have been demonstrated for complex biological samples originating from a sustained yeast cellular culture inside of a microfluidic device. The potential of the combination of microfluidics and IM-MS has been shown by the sheer number of analytes detected from a simple Jurkat cell population. This Jurkat cell data illustrates the abundance of secreted material which is observable with the use of ion mobility-mass spectrometry. Additionally, we have reviewed a study on cocaine metabolism in Jurkat cells which benchmarks the platform while also demonstrating its potential for discovery-driven research.
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