Capillary isoelectric focusing with pH 9.7 cathode for the analysis of gastric biopsies
Capillary isoelectric focusing tends to suffer from poor reproducibility, particularly for the analysis of complex protein samples from cellular or tissue homogenates. This poor reproducibility appears to be associated with erratic variations in electroosmotic flow. One cause of electroosmotic flow...
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| Veröffentlicht in: | Analytical and bioanalytical chemistry 2011-06, Vol.400 (7), p.2025-2030 |
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| creator | Ramsay, Lauren M. Cermak, Nathan Dada, Oluwatosin O. Dovichi, Norman J. |
| description | Capillary isoelectric focusing tends to suffer from poor reproducibility, particularly for the analysis of complex protein samples from cellular or tissue homogenates. This poor reproducibility appears to be associated with erratic variations in electroosmotic flow. One cause of electroosmotic flow variation is degradation of the capillary coating caused by the extremely basic solution commonly used during mobilization and focusing; this degradation of the capillary coating can be reduced by employing a CAPS mobilization buffer at pH 9. Another cause of variation is protein adsorption to the capillary wall, which causes an increase in electroosmotic flow. The effects of protein adsorption can be reduced by use of surfactants in the buffer and by employing an extremely low sample loading. We report the use of CAPS mobilization buffer in combination with an ultrasensitive laser-induced fluorescence detector for the reproducible analysis of ∼2 ng of protein from a Barrett's esophagus biopsy. |
| doi_str_mv | 10.1007/s00216-011-4926-2 |
| format | Article |
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This poor reproducibility appears to be associated with erratic variations in electroosmotic flow. One cause of electroosmotic flow variation is degradation of the capillary coating caused by the extremely basic solution commonly used during mobilization and focusing; this degradation of the capillary coating can be reduced by employing a CAPS mobilization buffer at pH 9. Another cause of variation is protein adsorption to the capillary wall, which causes an increase in electroosmotic flow. The effects of protein adsorption can be reduced by use of surfactants in the buffer and by employing an extremely low sample loading. We report the use of CAPS mobilization buffer in combination with an ultrasensitive laser-induced fluorescence detector for the reproducible analysis of ∼2 ng of protein from a Barrett's esophagus biopsy.</description><identifier>ISSN: 1618-2642</identifier><identifier>EISSN: 1618-2650</identifier><identifier>DOI: 10.1007/s00216-011-4926-2</identifier><identifier>PMID: 21461616</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer-Verlag</publisher><subject>Adsorption ; Analytical Chemistry ; Barrett Esophagus - pathology ; Biochemistry ; Biopsy ; Buffers ; Capillarity ; Characterization and Evaluation of Materials ; Chemistry ; Chemistry and Materials Science ; Coating ; Coatings ; Degradation ; Detectors ; Electrodes ; Exact sciences and technology ; Fluorescence ; Food Science ; Humans ; Hydrogen-Ion Concentration ; Isoelectric focusing ; Isoelectric Focusing - methods ; Laboratory Medicine ; Monitoring/Environmental Analysis ; Original Paper ; Protein adsorption ; Proteins - chemistry ; Reproducibility ; Reproducibility of Results ; Spectrometric and optical methods ; Spectrometry, Fluorescence ; Stomach - pathology ; Surface active agents ; Surface-Active Agents - chemistry</subject><ispartof>Analytical and bioanalytical chemistry, 2011-06, Vol.400 (7), p.2025-2030</ispartof><rights>Springer-Verlag 2011</rights><rights>2015 INIST-CNRS</rights><rights>COPYRIGHT 2011 Springer</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c520t-7bc479ccc08411e246167146283a6386cbd59c918ff3ad4198560600c8bf8f83</citedby><cites>FETCH-LOGICAL-c520t-7bc479ccc08411e246167146283a6386cbd59c918ff3ad4198560600c8bf8f83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00216-011-4926-2$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00216-011-4926-2$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,777,781,27905,27906,41469,42538,51300</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24235867$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21461616$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ramsay, Lauren M.</creatorcontrib><creatorcontrib>Cermak, Nathan</creatorcontrib><creatorcontrib>Dada, Oluwatosin O.</creatorcontrib><creatorcontrib>Dovichi, Norman J.</creatorcontrib><title>Capillary isoelectric focusing with pH 9.7 cathode for the analysis of gastric biopsies</title><title>Analytical and bioanalytical chemistry</title><addtitle>Anal Bioanal Chem</addtitle><addtitle>Anal Bioanal Chem</addtitle><description>Capillary isoelectric focusing tends to suffer from poor reproducibility, particularly for the analysis of complex protein samples from cellular or tissue homogenates. This poor reproducibility appears to be associated with erratic variations in electroosmotic flow. One cause of electroosmotic flow variation is degradation of the capillary coating caused by the extremely basic solution commonly used during mobilization and focusing; this degradation of the capillary coating can be reduced by employing a CAPS mobilization buffer at pH 9. Another cause of variation is protein adsorption to the capillary wall, which causes an increase in electroosmotic flow. The effects of protein adsorption can be reduced by use of surfactants in the buffer and by employing an extremely low sample loading. We report the use of CAPS mobilization buffer in combination with an ultrasensitive laser-induced fluorescence detector for the reproducible analysis of ∼2 ng of protein from a Barrett's esophagus biopsy.</description><subject>Adsorption</subject><subject>Analytical Chemistry</subject><subject>Barrett Esophagus - pathology</subject><subject>Biochemistry</subject><subject>Biopsy</subject><subject>Buffers</subject><subject>Capillarity</subject><subject>Characterization and Evaluation of Materials</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Coating</subject><subject>Coatings</subject><subject>Degradation</subject><subject>Detectors</subject><subject>Electrodes</subject><subject>Exact sciences and technology</subject><subject>Fluorescence</subject><subject>Food Science</subject><subject>Humans</subject><subject>Hydrogen-Ion Concentration</subject><subject>Isoelectric focusing</subject><subject>Isoelectric Focusing - methods</subject><subject>Laboratory Medicine</subject><subject>Monitoring/Environmental Analysis</subject><subject>Original Paper</subject><subject>Protein adsorption</subject><subject>Proteins - chemistry</subject><subject>Reproducibility</subject><subject>Reproducibility of Results</subject><subject>Spectrometric and optical methods</subject><subject>Spectrometry, Fluorescence</subject><subject>Stomach - pathology</subject><subject>Surface active agents</subject><subject>Surface-Active Agents - chemistry</subject><issn>1618-2642</issn><issn>1618-2650</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcFu1DAURSMEoqXwAWxQNqhsMrznOI69rEaUIlVi0wU7y3HsGVeZOPglQv0bvoUvw0OGsivywpbvufa1b1G8RdggQPuRABiKChArrpio2LPiHAXKiokGnj-uOTsrXhHdA2AjUbwszhhykUVxXnzbmikMg0kPZaDoBmfnFGzpo10ojLvyR5j35XTz66fatKU18z72LqupnPeuNKMZHihQGX25M_TH2YU4UXD0unjhzUDuzWm-KO6uP91tb6rbr5-_bK9uK9swmKu2s7xV1lqQHNGxY642p2OyNqKWwnZ9o6xC6X1teo5KNgIEgJWdl17WF8XleuyU4vfF0awPgazLLxpdXEgrVKpuGfyflEJCDUJBJj88SWLLeQtY15jRzYruzOB0GH2ck7F59O4QbBydD3n_qlYNZ6CaJhtwNdgUiZLzekrhkL9fI-hjqXotVedS9bFUzbLn3SnP0h1c_-j422IG3p8AQ9YMPpnRBvrHcVY3UrSZYytHWRp3Lun7uKRcIj1x-29biLeY</recordid><startdate>20110601</startdate><enddate>20110601</enddate><creator>Ramsay, Lauren M.</creator><creator>Cermak, Nathan</creator><creator>Dada, Oluwatosin O.</creator><creator>Dovichi, Norman J.</creator><general>Springer-Verlag</general><general>Springer</general><scope>IQODW</scope><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>7SR</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>L7M</scope><scope>7X8</scope><scope>7QH</scope><scope>7UA</scope><scope>C1K</scope></search><sort><creationdate>20110601</creationdate><title>Capillary isoelectric focusing with pH 9.7 cathode for the analysis of gastric biopsies</title><author>Ramsay, Lauren M. ; Cermak, Nathan ; Dada, Oluwatosin O. ; Dovichi, Norman J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c520t-7bc479ccc08411e246167146283a6386cbd59c918ff3ad4198560600c8bf8f83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Adsorption</topic><topic>Analytical Chemistry</topic><topic>Barrett Esophagus - pathology</topic><topic>Biochemistry</topic><topic>Biopsy</topic><topic>Buffers</topic><topic>Capillarity</topic><topic>Characterization and Evaluation of Materials</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Coating</topic><topic>Coatings</topic><topic>Degradation</topic><topic>Detectors</topic><topic>Electrodes</topic><topic>Exact sciences and technology</topic><topic>Fluorescence</topic><topic>Food Science</topic><topic>Humans</topic><topic>Hydrogen-Ion Concentration</topic><topic>Isoelectric focusing</topic><topic>Isoelectric Focusing - methods</topic><topic>Laboratory Medicine</topic><topic>Monitoring/Environmental Analysis</topic><topic>Original Paper</topic><topic>Protein adsorption</topic><topic>Proteins - chemistry</topic><topic>Reproducibility</topic><topic>Reproducibility of Results</topic><topic>Spectrometric and optical methods</topic><topic>Spectrometry, Fluorescence</topic><topic>Stomach - pathology</topic><topic>Surface active agents</topic><topic>Surface-Active Agents - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ramsay, Lauren M.</creatorcontrib><creatorcontrib>Cermak, Nathan</creatorcontrib><creatorcontrib>Dada, Oluwatosin O.</creatorcontrib><creatorcontrib>Dovichi, Norman J.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Engineered Materials Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>MEDLINE - Academic</collection><collection>Aqualine</collection><collection>Water Resources Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Analytical and bioanalytical chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ramsay, Lauren M.</au><au>Cermak, Nathan</au><au>Dada, Oluwatosin O.</au><au>Dovichi, Norman J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Capillary isoelectric focusing with pH 9.7 cathode for the analysis of gastric biopsies</atitle><jtitle>Analytical and bioanalytical chemistry</jtitle><stitle>Anal Bioanal Chem</stitle><addtitle>Anal Bioanal Chem</addtitle><date>2011-06-01</date><risdate>2011</risdate><volume>400</volume><issue>7</issue><spage>2025</spage><epage>2030</epage><pages>2025-2030</pages><issn>1618-2642</issn><eissn>1618-2650</eissn><abstract>Capillary isoelectric focusing tends to suffer from poor reproducibility, particularly for the analysis of complex protein samples from cellular or tissue homogenates. This poor reproducibility appears to be associated with erratic variations in electroosmotic flow. One cause of electroosmotic flow variation is degradation of the capillary coating caused by the extremely basic solution commonly used during mobilization and focusing; this degradation of the capillary coating can be reduced by employing a CAPS mobilization buffer at pH 9. Another cause of variation is protein adsorption to the capillary wall, which causes an increase in electroosmotic flow. The effects of protein adsorption can be reduced by use of surfactants in the buffer and by employing an extremely low sample loading. 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| subjects | Adsorption Analytical Chemistry Barrett Esophagus - pathology Biochemistry Biopsy Buffers Capillarity Characterization and Evaluation of Materials Chemistry Chemistry and Materials Science Coating Coatings Degradation Detectors Electrodes Exact sciences and technology Fluorescence Food Science Humans Hydrogen-Ion Concentration Isoelectric focusing Isoelectric Focusing - methods Laboratory Medicine Monitoring/Environmental Analysis Original Paper Protein adsorption Proteins - chemistry Reproducibility Reproducibility of Results Spectrometric and optical methods Spectrometry, Fluorescence Stomach - pathology Surface active agents Surface-Active Agents - chemistry |
| title | Capillary isoelectric focusing with pH 9.7 cathode for the analysis of gastric biopsies |
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