Advanced cleanup process of the free-flow microfluidic device for protein analysis
The treatment of samples preparation is generally recognized as a bottleneck for the rapid analysis of protein because of the off-chip performance in many cases. In this study, we used the charge characteristics of protein to develop a simple and rapid electro-microfluidic desalting system as an eff...
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| Veröffentlicht in: | Ultramicroscopy 2008-09, Vol.108 (10), p.1365-1370 |
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| container_title | Ultramicroscopy |
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| creator | Huh, Yun Suk Park, Tae Jung Yang, KwangSuk Lee, Eun Zoo Hong, Yeon Ki Lee, Sang Yup Kim, Do Hyun Hong, Won Hi |
| description | The treatment of samples preparation is generally recognized as a bottleneck for the rapid analysis of protein because of the off-chip performance in many cases. In this study, we used the charge characteristics of protein to develop a simple and rapid electro-microfluidic desalting system as an effective means of cleaning up protein sample. When we loaded a urea-rich protein sample and a buffer solution into a free-flow zone electrophoresis (FFZE) chamber, the microfluidic device was able to separate the charged protein sample and the non-charged urea. With a 90
V electric field in the FFZE chamber, the removal efficiency of the urea was about 88% and the recovery of the protein was 78%. In addition, the desalted protein sample used in this device showed significant improvement with respect to the MALDI-TOF-MS spectrum signal of a fusion protein, which was fused to the gold-binding polypeptide with enhanced green fluorescent protein, as a model protein. The inflow of the purified fusion protein sample can be successfully immobilized on the gold surface and analyzed by confocal fluorescence microscopy and surface plasmon resonance for biotechnological sensors. |
| doi_str_mv | 10.1016/j.ultramic.2008.04.085 |
| format | Article |
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V electric field in the FFZE chamber, the removal efficiency of the urea was about 88% and the recovery of the protein was 78%. In addition, the desalted protein sample used in this device showed significant improvement with respect to the MALDI-TOF-MS spectrum signal of a fusion protein, which was fused to the gold-binding polypeptide with enhanced green fluorescent protein, as a model protein. The inflow of the purified fusion protein sample can be successfully immobilized on the gold surface and analyzed by confocal fluorescence microscopy and surface plasmon resonance for biotechnological sensors.</description><identifier>ISSN: 0304-3991</identifier><identifier>EISSN: 1879-2723</identifier><identifier>DOI: 10.1016/j.ultramic.2008.04.085</identifier><identifier>PMID: 18562114</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Biosensing Techniques ; Biosensor ; Electro-desalting ; Electrophoresis - instrumentation ; Electrophoresis - methods ; Fusion protein ; Green Fluorescent Proteins - analysis ; Green Fluorescent Proteins - chemistry ; Green Fluorescent Proteins - genetics ; Green Fluorescent Proteins - metabolism ; Microfluidic Analytical Techniques - instrumentation ; Microfluidic Analytical Techniques - methods ; Microfluidic device ; Microfluidics - instrumentation ; Microfluidics - methods ; Proteins - analysis ; Proteins - chemistry ; Recombinant Fusion Proteins - analysis ; Recombinant Fusion Proteins - chemistry ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - metabolism ; Sample preparation ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods ; Urea - chemistry</subject><ispartof>Ultramicroscopy, 2008-09, Vol.108 (10), p.1365-1370</ispartof><rights>2008 Elsevier B.V.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c397t-beaf86b9ce1252e1718ced1b09bfebb42a6cc206e932326ba7ea3aa00ae3f0223</citedby><cites>FETCH-LOGICAL-c397t-beaf86b9ce1252e1718ced1b09bfebb42a6cc206e932326ba7ea3aa00ae3f0223</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.ultramic.2008.04.085$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18562114$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Huh, Yun Suk</creatorcontrib><creatorcontrib>Park, Tae Jung</creatorcontrib><creatorcontrib>Yang, KwangSuk</creatorcontrib><creatorcontrib>Lee, Eun Zoo</creatorcontrib><creatorcontrib>Hong, Yeon Ki</creatorcontrib><creatorcontrib>Lee, Sang Yup</creatorcontrib><creatorcontrib>Kim, Do Hyun</creatorcontrib><creatorcontrib>Hong, Won Hi</creatorcontrib><title>Advanced cleanup process of the free-flow microfluidic device for protein analysis</title><title>Ultramicroscopy</title><addtitle>Ultramicroscopy</addtitle><description>The treatment of samples preparation is generally recognized as a bottleneck for the rapid analysis of protein because of the off-chip performance in many cases. In this study, we used the charge characteristics of protein to develop a simple and rapid electro-microfluidic desalting system as an effective means of cleaning up protein sample. When we loaded a urea-rich protein sample and a buffer solution into a free-flow zone electrophoresis (FFZE) chamber, the microfluidic device was able to separate the charged protein sample and the non-charged urea. With a 90
V electric field in the FFZE chamber, the removal efficiency of the urea was about 88% and the recovery of the protein was 78%. In addition, the desalted protein sample used in this device showed significant improvement with respect to the MALDI-TOF-MS spectrum signal of a fusion protein, which was fused to the gold-binding polypeptide with enhanced green fluorescent protein, as a model protein. The inflow of the purified fusion protein sample can be successfully immobilized on the gold surface and analyzed by confocal fluorescence microscopy and surface plasmon resonance for biotechnological sensors.</description><subject>Animals</subject><subject>Biosensing Techniques</subject><subject>Biosensor</subject><subject>Electro-desalting</subject><subject>Electrophoresis - instrumentation</subject><subject>Electrophoresis - methods</subject><subject>Fusion protein</subject><subject>Green Fluorescent Proteins - analysis</subject><subject>Green Fluorescent Proteins - chemistry</subject><subject>Green Fluorescent Proteins - genetics</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Microfluidic Analytical Techniques - instrumentation</subject><subject>Microfluidic Analytical Techniques - methods</subject><subject>Microfluidic device</subject><subject>Microfluidics - instrumentation</subject><subject>Microfluidics - methods</subject><subject>Proteins - analysis</subject><subject>Proteins - chemistry</subject><subject>Recombinant Fusion Proteins - analysis</subject><subject>Recombinant Fusion Proteins - chemistry</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Sample preparation</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</subject><subject>Urea - chemistry</subject><issn>0304-3991</issn><issn>1879-2723</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMtOwzAQRS0EouXxCygrdgljO3XsHQjxkiohIVhbjjMWrtKk2EkRf4-jFrHsahZzru_4EHJFoaBAxc2qGNshmLW3BQOQBZQFyMURmVNZqZxVjB-TOXAoc64UnZGzGFcAQKGUp2RG5UIwSss5ebtrtqaz2GS2RdONm2wTeosxZr3Lhk_MXEDMXdt_Z6kr9K4dfeNt1uDW27TtwxQY0HeZ6Uz7E328ICfOtBEv9_OcfDw-vN8_58vXp5f7u2VuuaqGvEbjpKiVRcoWDGlFZTqD1qBqh3VdMiOsZSBQccaZqE2FhhsDYJA7YIyfk-vdu-mArxHjoNc-Wmxb02E_Ri1UKYXicBBkoKhUJU2g2IHpozEGdHoT_NqEH01BT9r1Sv9p15N2DaVO2lPwat8w1mts_mN7zwm43QGYhGw9Bh2tx8m7D2gH3fT-UMcvOWmYzg</recordid><startdate>20080901</startdate><enddate>20080901</enddate><creator>Huh, Yun Suk</creator><creator>Park, Tae Jung</creator><creator>Yang, KwangSuk</creator><creator>Lee, Eun Zoo</creator><creator>Hong, Yeon Ki</creator><creator>Lee, Sang Yup</creator><creator>Kim, Do Hyun</creator><creator>Hong, Won Hi</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20080901</creationdate><title>Advanced cleanup process of the free-flow microfluidic device for protein analysis</title><author>Huh, Yun Suk ; Park, Tae Jung ; Yang, KwangSuk ; Lee, Eun Zoo ; Hong, Yeon Ki ; Lee, Sang Yup ; Kim, Do Hyun ; Hong, Won Hi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c397t-beaf86b9ce1252e1718ced1b09bfebb42a6cc206e932326ba7ea3aa00ae3f0223</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Animals</topic><topic>Biosensing Techniques</topic><topic>Biosensor</topic><topic>Electro-desalting</topic><topic>Electrophoresis - instrumentation</topic><topic>Electrophoresis - methods</topic><topic>Fusion protein</topic><topic>Green Fluorescent Proteins - analysis</topic><topic>Green Fluorescent Proteins - chemistry</topic><topic>Green Fluorescent Proteins - genetics</topic><topic>Green Fluorescent Proteins - metabolism</topic><topic>Microfluidic Analytical Techniques - instrumentation</topic><topic>Microfluidic Analytical Techniques - methods</topic><topic>Microfluidic device</topic><topic>Microfluidics - instrumentation</topic><topic>Microfluidics - methods</topic><topic>Proteins - analysis</topic><topic>Proteins - chemistry</topic><topic>Recombinant Fusion Proteins - analysis</topic><topic>Recombinant Fusion Proteins - chemistry</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Sample preparation</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</topic><topic>Urea - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Huh, Yun Suk</creatorcontrib><creatorcontrib>Park, Tae Jung</creatorcontrib><creatorcontrib>Yang, KwangSuk</creatorcontrib><creatorcontrib>Lee, Eun Zoo</creatorcontrib><creatorcontrib>Hong, Yeon Ki</creatorcontrib><creatorcontrib>Lee, Sang Yup</creatorcontrib><creatorcontrib>Kim, Do Hyun</creatorcontrib><creatorcontrib>Hong, Won Hi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Ultramicroscopy</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Huh, Yun Suk</au><au>Park, Tae Jung</au><au>Yang, KwangSuk</au><au>Lee, Eun Zoo</au><au>Hong, Yeon Ki</au><au>Lee, Sang Yup</au><au>Kim, Do Hyun</au><au>Hong, Won Hi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Advanced cleanup process of the free-flow microfluidic device for protein analysis</atitle><jtitle>Ultramicroscopy</jtitle><addtitle>Ultramicroscopy</addtitle><date>2008-09-01</date><risdate>2008</risdate><volume>108</volume><issue>10</issue><spage>1365</spage><epage>1370</epage><pages>1365-1370</pages><issn>0304-3991</issn><eissn>1879-2723</eissn><abstract>The treatment of samples preparation is generally recognized as a bottleneck for the rapid analysis of protein because of the off-chip performance in many cases. 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V electric field in the FFZE chamber, the removal efficiency of the urea was about 88% and the recovery of the protein was 78%. In addition, the desalted protein sample used in this device showed significant improvement with respect to the MALDI-TOF-MS spectrum signal of a fusion protein, which was fused to the gold-binding polypeptide with enhanced green fluorescent protein, as a model protein. The inflow of the purified fusion protein sample can be successfully immobilized on the gold surface and analyzed by confocal fluorescence microscopy and surface plasmon resonance for biotechnological sensors.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>18562114</pmid><doi>10.1016/j.ultramic.2008.04.085</doi><tpages>6</tpages></addata></record> |
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| subjects | Animals Biosensing Techniques Biosensor Electro-desalting Electrophoresis - instrumentation Electrophoresis - methods Fusion protein Green Fluorescent Proteins - analysis Green Fluorescent Proteins - chemistry Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism Microfluidic Analytical Techniques - instrumentation Microfluidic Analytical Techniques - methods Microfluidic device Microfluidics - instrumentation Microfluidics - methods Proteins - analysis Proteins - chemistry Recombinant Fusion Proteins - analysis Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Sample preparation Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods Urea - chemistry |
| title | Advanced cleanup process of the free-flow microfluidic device for protein analysis |
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