Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry
In this paper, we are evaluating the strategy of sorting peptides/proteins based on the charge to mass without resorting to ampholytes and/or isoelectric focusing, using a single- and two-step free-flow zone electrophoresis. We developed a simple fabrication method to create a salt bridge for free-f...
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| Format: | Article |
| Language: | en_US |
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American Chemical Society
2011
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| Online Access: | http://hdl.handle.net/1721.1/66895 https://orcid.org/0000-0002-2325-552X https://orcid.org/0000-0001-7215-1439 |
| _version_ | 1826204558663090176 |
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| author | Chan, Michael Song, Yong-Ak Celio, Chris Tannenbaum, Steven Robert Wishnok, John S. Han, Jongyoon |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Chan, Michael Song, Yong-Ak Celio, Chris Tannenbaum, Steven Robert Wishnok, John S. Han, Jongyoon |
| author_sort | Chan, Michael |
| collection | MIT |
| description | In this paper, we are evaluating the strategy of sorting peptides/proteins based on the charge to mass without resorting to ampholytes and/or isoelectric focusing, using a single- and two-step free-flow zone electrophoresis. We developed a simple fabrication method to create a salt bridge for free-flow zone electrophoresis in PDMS chips by surface printing a hydrophobic layer on a glass substrate. Since the surface-printed hydrophobic layer prevents plasma bonding between the PDMS chip and the substrate, an electrical junction gap can be created for free-flow zone electrophoresis. With this device, we demonstrated a separation of positive and negative peptides and proteins at a given pH in standard buffer systems and validated the sorting result with LC/MS. Furthermore, we coupled two sorting steps via off-chip titration and isolated peptides within specific pI ranges from sample mixtures, where the pI range was simply set by the pH values of the buffer solutions. This free-flow zone electrophoresis sorting device, with its simplicity of fabrication, and a sorting resolution of 0.5 pH unit, can potentially be a high-throughput sample fractionation tool for targeted proteomics using LC/MS. |
| first_indexed | 2024-09-23T12:57:22Z |
| format | Article |
| id | mit-1721.1/66895 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T12:57:22Z |
| publishDate | 2011 |
| publisher | American Chemical Society |
| record_format | dspace |
| spelling | mit-1721.1/668952022-10-01T12:06:03Z Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry Chan, Michael Song, Yong-Ak Celio, Chris Tannenbaum, Steven Robert Wishnok, John S. Han, Jongyoon Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Chemistry Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Massachusetts Institute of Technology. Research Laboratory of Electronics Tannenbaum, Steven R. Song, Yong-Ak Celio, Chris Tannenbaum, Steven Robert Wishnok, John S. Han, Jongyoon In this paper, we are evaluating the strategy of sorting peptides/proteins based on the charge to mass without resorting to ampholytes and/or isoelectric focusing, using a single- and two-step free-flow zone electrophoresis. We developed a simple fabrication method to create a salt bridge for free-flow zone electrophoresis in PDMS chips by surface printing a hydrophobic layer on a glass substrate. Since the surface-printed hydrophobic layer prevents plasma bonding between the PDMS chip and the substrate, an electrical junction gap can be created for free-flow zone electrophoresis. With this device, we demonstrated a separation of positive and negative peptides and proteins at a given pH in standard buffer systems and validated the sorting result with LC/MS. Furthermore, we coupled two sorting steps via off-chip titration and isolated peptides within specific pI ranges from sample mixtures, where the pI range was simply set by the pH values of the buffer solutions. This free-flow zone electrophoresis sorting device, with its simplicity of fabrication, and a sorting resolution of 0.5 pH unit, can potentially be a high-throughput sample fractionation tool for targeted proteomics using LC/MS. Korea Institute of Science and Technology. Intelligent Microsystems Center Massachusetts Institute of Technology. Center for Environmental Health Sciences National Institute of Environmental Health Sciences (Grant No. P30-ES002109) United States. National Institutes of Health (grant R21 EB008177) 2011-11-02T19:47:12Z 2011-11-02T19:47:12Z 2010-02 Article http://purl.org/eprint/type/JournalArticle 0003-2700 1520-6882 http://hdl.handle.net/1721.1/66895 Song, Yong-Ak et al. “Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry.” Analytical Chemistry 82 (2010): 2317-2325. Web. 2 Nov. 2011. © 2011 American Chemical Society https://orcid.org/0000-0002-2325-552X https://orcid.org/0000-0001-7215-1439 en_US http://dx.doi.org/10.1021/ac9025219 Analytical Chemistry Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf American Chemical Society PubMed Central |
| spellingShingle | Chan, Michael Song, Yong-Ak Celio, Chris Tannenbaum, Steven Robert Wishnok, John S. Han, Jongyoon Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry |
| title | Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry |
| title_full | Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry |
| title_fullStr | Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry |
| title_full_unstemmed | Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry |
| title_short | Free-Flow Zone Electrophoresis of Peptides and Proteins in PDMS Microchip for Narrow pI Range Sample Prefractionation Coupled with Mass Spectrometry |
| title_sort | free flow zone electrophoresis of peptides and proteins in pdms microchip for narrow pi range sample prefractionation coupled with mass spectrometry |
| url | http://hdl.handle.net/1721.1/66895 https://orcid.org/0000-0002-2325-552X https://orcid.org/0000-0001-7215-1439 |
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