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A Plug-Based Microfluidic System for Dispensing Lipidic Cubic Phase (LCP) Material Validated by Crystallizing
Liang Li1, Qiang Fu, Christopher A Kors
1Department of Chemistry and Institute for Biophysical Dynamics The University of Chicago, 929 East 57 Street, Chicago, IL, 60637.
Microfluidics and Nanofluidics
|May 18, 2010
Summary
A novel microfluidic system accurately dispenses nanoliter Lipidic Cubic Phase (LCP) plugs, preventing dehydration and enabling membrane protein crystallization. This method, including Post LCP-formation Incorporation (PLI), minimizes sample use and expands screening capabilities.
Area of Science:
- Biophysics
- Structural Biology
- Microfluidics
Background:
- Membrane proteins are crucial for cellular functions but challenging to crystallize.
- Lipidic Cubic Phase (LCP) is a promising matrix for membrane protein crystallization.
- Traditional LCP methods face challenges with sample volume and dehydration.
Purpose of the Study:
- To develop a plug-based microfluidic system for precise nanoliter LCP dispensing.
- To facilitate membrane protein crystallization using LCP.
- To introduce and validate the Post LCP-formation Incorporation (PLI) method.
Main Methods:
- A microfluidic system was designed to dispense and merge nanoliter LCP and aqueous plugs.
- LCP plug formation conditions were characterized and presented in a phase diagram.
- Membrane proteins were incorporated using both traditional methods and PLI.
Main Results:
- The system successfully crystallized bacteriorhodopsin using traditional LCP formation.
- Crystals of bacterial photosynthetic reaction centers were obtained using the PLI method.
- The microfluidic system demonstrated accurate LCP dispensing and prevented dehydration.
Conclusions:
- The plug-based microfluidic system offers precise control over LCP dispensing for membrane protein crystallization.
- The PLI method combined with microfluidics reduces sample consumption and broadens screening parameters.
- This approach is valuable for advancing membrane protein structure determination.
