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Microfluidics-based strategies for protein crystallography

Citation

Anderson, Megan J. (2009) Microfluidics-based strategies for protein crystallography. Dissertation (Ph.D.), California Institute of Technology. http://resolver.caltech.edu/CaltechETD:etd-10172008-222221

Abstract

Protein crystallography is an invaluable tool for the study of biological processes at the molecular level. While several crystallization techniques are actively pursued in both academic and industrial laboratories to produce high-quality protein crystals, the use of microfluidic technology for structural biology was previously shown to improve protein crystallization over more traditional methods. This thesis describes a microfluidics-based crystallization strategy that was developed to increase the success rate of crystallizing challenging proteins. The crystallization strategy involves using multiple microfluidic devices to characterize the solubility trends of the crystallization target, to perform nanoliter volume free interface diffusion crystallization experiments designed around the solubility trends, and to enable in situ diffraction analysis of crystals grown in microfluidic devices. The crystallization strategy was applied to the crystallization of a dozen challenging proteins and increased the overall crystallization and diffraction success rates compared with conventional automation. The crystallization strategy was also utilized to crystallize four metabolic proteins and provides the first demonstration of in situ structure determination for novel crystallization targets using a microfluidic crystallization platform. Additional technological advances were accomplished by the development of a novel microfluidic device designed to address the specific challenges of membrane protein crystallography. To date, this microfluidic crystallization strategy has produced four novel protein structures and holds great promise for future work in the field of protein crystallography.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:crystallography; microfluidics
Degree Grantor:California Institute of Technology
Division:Biology
Major Option:Biochemistry and Molecular Biophysics
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Quake, Stephen R.
Thesis Committee:
  • Quake, Stephen R. (chair)
  • Rees, Douglas C. (co-chair)
  • Bjorkman, Pamela J.
  • Mayo, Stephen L.
Defense Date:18 September 2008
Record Number:CaltechETD:etd-10172008-222221
Persistent URL:http://resolver.caltech.edu/CaltechETD:etd-10172008-222221
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:4147
Collection:CaltechTHESIS
Deposited By: Imported from ETD-db
Deposited On:11 Nov 2008
Last Modified:06 Jan 2014 18:27

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