Microfluidic preparative free-flow isoelectric focusing in a triangular channel: System development and characterization
Name
Jensen_Microfluidic preparative.pdf
Size
1.53 MB
Format
Adobe PDF
Checksum (MD5)
dd0e21220b7e3f03b611e7d98f7cc175
Author(s)
Wen, Jian
Albrecht, Jacob W.
Jensen, Klavs F.
Date Issued
April 2010
April 2010
Journal
ELECTROPHORESIS
Publisher
John Wiley & Sons, Inc.
Citation
Wen, Jian, Jacob W. Albrecht, and Klavs F. Jensen. Microfluidic Preparative Free-flow Isoelectric Focusing in a Triangular Channel: System Development and Characterization. Electrophoresis 31, no. 10 (May 23, 2010): 1606-1614.
Version
Author's final manuscript
Abstract
A preparative scale free-flow IEF device is developed and characterized with the aim of addressing needs of molecular biologists working with protein samples on the milligrams and milliliters scale. A triangular-shape separation channel facilitates the establishment of the pH gradient with a corresponding increase in separation efficiency and decrease in focusing time compared with that in a regular rectangular channel. Functionalized, ion-permeable poly(acrylamide) gel membranes are sandwiched between PDMS and glass layers to both isolate the electrode buffers from the central separation channel and also to selectively adjust the voltage efficiency across the separation channel to achieve high electric field separation. The 50×70 mm device is fabricated by soft lithography and has 24 outlets evenly spaced across a pH gradient between pH 4 and 10. This preparative free-flow IEF system is investigated and optimized for both aqueous and denaturing conditions with respect to the electric field and potential efficiency and with consideration of Joule-heating removal. Energy distribution across the functionalized polyacrylamide gel is investigated and controlled to adjust the potential efficiency between 15 and 80% across the triangular separation channel. The device is able to achieve constant electric fields high as 370±20 V/cm through the entire triangular channel given the separation voltage of 1800 V, enabling separation of five fluorescent pI markers as a demonstration example.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Terms of Use
Creative Commons Attribution-Noncommercial-Share Alike 3.0
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1002/elps.200900577
