Microfluidic preparative free-flow isoelectric focusing in a triangular channel: System development and characterization
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 corresp...
| Main Authors: | , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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John Wiley & Sons, Inc.
2013
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| Online Access: | http://hdl.handle.net/1721.1/79374 https://orcid.org/0000-0001-7192-580X |
| _version_ | 1811080115773243392 |
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| author | Wen, Jian Albrecht, Jacob W. Jensen, Klavs F. |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Wen, Jian Albrecht, Jacob W. Jensen, Klavs F. |
| author_sort | Wen, Jian |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T11:26:04Z |
| format | Article |
| id | mit-1721.1/79374 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:26:04Z |
| publishDate | 2013 |
| publisher | John Wiley & Sons, Inc. |
| record_format | dspace |
| spelling | mit-1721.1/793742022-10-01T03:34:45Z Microfluidic preparative free-flow isoelectric focusing in a triangular channel: System development and characterization Wen, Jian Albrecht, Jacob W. Jensen, Klavs F. Massachusetts Institute of Technology. Department of Chemical Engineering Jensen, Klavs F. 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. United States. Army Research Office (Grant Number: W911NF-07-D-0004) National Institutes of Health (U.S.) (Grant Number: GM68762) 2013-06-26T16:01:32Z 2013-06-26T16:01:32Z 2010-04 2010-04 Article http://purl.org/eprint/type/JournalArticle 01730835 15222683 http://hdl.handle.net/1721.1/79374 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. https://orcid.org/0000-0001-7192-580X en_US http://dx.doi.org/10.1002/elps.200900577 ELECTROPHORESIS Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf John Wiley & Sons, Inc. PMC |
| spellingShingle | Wen, Jian Albrecht, Jacob W. Jensen, Klavs F. Microfluidic preparative free-flow isoelectric focusing in a triangular channel: System development and characterization |
| title | Microfluidic preparative free-flow isoelectric focusing in a triangular channel: System development and characterization |
| title_full | Microfluidic preparative free-flow isoelectric focusing in a triangular channel: System development and characterization |
| title_fullStr | Microfluidic preparative free-flow isoelectric focusing in a triangular channel: System development and characterization |
| title_full_unstemmed | Microfluidic preparative free-flow isoelectric focusing in a triangular channel: System development and characterization |
| title_short | Microfluidic preparative free-flow isoelectric focusing in a triangular channel: System development and characterization |
| title_sort | microfluidic preparative free flow isoelectric focusing in a triangular channel system development and characterization |
| url | http://hdl.handle.net/1721.1/79374 https://orcid.org/0000-0001-7192-580X |
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