Development of a sandwich ELISA for the 5.9-kDa fibrinogen alpha C chain fragment detected by serum proteome analysis
Purpose: We previously identified novel biomarker candidates in heavy consumers of alcohol using serum proteome analysis. Among several candidates, a 5.9 kDa peptide identified as a fragment of the fibrinogen alpha C chain (FIC5.9) was the most promising. To move FIC5.9 toward potential diagnostic u...
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| Veröffentlicht in: | Proteomics. Clinical applications 2011-04, Vol.5 (3-4), p.141-146 |
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| container_title | Proteomics. Clinical applications |
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| creator | Noda, Kenta Sogawa, Kazuyuki Kikuchi, Wataru Kiyokawa, Iwao Miura, Toshihide Kojima, Ryo Katayama, Katsuhiro Kodera, Yoshio Nomura, Fumio |
| description | Purpose: We previously identified novel biomarker candidates in heavy consumers of alcohol using serum proteome analysis. Among several candidates, a 5.9 kDa peptide identified as a fragment of the fibrinogen alpha C chain (FIC5.9) was the most promising. To move FIC5.9 toward potential diagnostic use, we developed an enzyme immunoassay that enables measurement of serum FIC5.9 levels.
Experimental design: Two monoclonal antibodies specific to the N and C‐termini of the 5.9‐kDa peptide were used to develop a FIC5.9 sandwich ELISA. The assay was evaluated by comparing the results with those obtained by the stable isotope‐labeled dilution mass spectrometry (SID‐MS) using the ClinProt™ system.
Results: The ELISA results correlated with the SID‐MS findings (slope=0.795, intercept=−0.011, r2=0.908) and the performance of the ELISA was satisfactory in terms of recovery (98.5–103.0%) and within‐run (1.4–4.7%) and between‐day (2.8–8.4%) reproducibility. The assay was capable of detecting changes in FIC5.9 during abstinence from drinking in patients with alcohol dependency (p |
| doi_str_mv | 10.1002/prca.201000127 |
| format | Article |
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Experimental design: Two monoclonal antibodies specific to the N and C‐termini of the 5.9‐kDa peptide were used to develop a FIC5.9 sandwich ELISA. The assay was evaluated by comparing the results with those obtained by the stable isotope‐labeled dilution mass spectrometry (SID‐MS) using the ClinProt™ system.
Results: The ELISA results correlated with the SID‐MS findings (slope=0.795, intercept=−0.011, r2=0.908) and the performance of the ELISA was satisfactory in terms of recovery (98.5–103.0%) and within‐run (1.4–4.7%) and between‐day (2.8–8.4%) reproducibility. The assay was capable of detecting changes in FIC5.9 during abstinence from drinking in patients with alcohol dependency (p<0.0001).
Conclusions and clinical relevance: The sandwich ELISA developed in this study will be useful for validation of the diagnostic significance of serum FIC5.9 levels in various pathological conditions, including alcoholism.</description><identifier>ISSN: 1862-8346</identifier><identifier>EISSN: 1862-8354</identifier><identifier>DOI: 10.1002/prca.201000127</identifier><identifier>PMID: 21360827</identifier><language>eng</language><publisher>Weinheim: WILEY-VCH Verlag</publisher><subject>Alcoholism - blood ; Biological and medical sciences ; Diverse techniques ; Enzyme-Linked Immunosorbent Assay - methods ; Fibrinogen ; Fibrinogen alpha C chain ; Fundamental and applied biological sciences. Psychology ; Humans ; Liver disease ; Male ; Mass Spectrometry ; Medical research ; Middle Aged ; Molecular and cellular biology ; Peptide Fragments - blood ; Proteome - analysis ; Proteomics ; Reproducibility of Results ; Sandwich ELISA ; Sensitivity and Specificity ; Serum biomarker ; Stable isotope-labeled internal standard dilution-MS (SID-MS)</subject><ispartof>Proteomics. Clinical applications, 2011-04, Vol.5 (3-4), p.141-146</ispartof><rights>Copyright © 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.</rights><rights>Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5707-8e29020fe5ae09a9c61254f96355b5b7273d39426b7dc7c460c57582978ea1053</citedby><cites>FETCH-LOGICAL-c5707-8e29020fe5ae09a9c61254f96355b5b7273d39426b7dc7c460c57582978ea1053</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fprca.201000127$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fprca.201000127$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24066954$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21360827$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Noda, Kenta</creatorcontrib><creatorcontrib>Sogawa, Kazuyuki</creatorcontrib><creatorcontrib>Kikuchi, Wataru</creatorcontrib><creatorcontrib>Kiyokawa, Iwao</creatorcontrib><creatorcontrib>Miura, Toshihide</creatorcontrib><creatorcontrib>Kojima, Ryo</creatorcontrib><creatorcontrib>Katayama, Katsuhiro</creatorcontrib><creatorcontrib>Kodera, Yoshio</creatorcontrib><creatorcontrib>Nomura, Fumio</creatorcontrib><title>Development of a sandwich ELISA for the 5.9-kDa fibrinogen alpha C chain fragment detected by serum proteome analysis</title><title>Proteomics. Clinical applications</title><addtitle>Prot. Clin. Appl</addtitle><description>Purpose: We previously identified novel biomarker candidates in heavy consumers of alcohol using serum proteome analysis. Among several candidates, a 5.9 kDa peptide identified as a fragment of the fibrinogen alpha C chain (FIC5.9) was the most promising. To move FIC5.9 toward potential diagnostic use, we developed an enzyme immunoassay that enables measurement of serum FIC5.9 levels.
Experimental design: Two monoclonal antibodies specific to the N and C‐termini of the 5.9‐kDa peptide were used to develop a FIC5.9 sandwich ELISA. The assay was evaluated by comparing the results with those obtained by the stable isotope‐labeled dilution mass spectrometry (SID‐MS) using the ClinProt™ system.
Results: The ELISA results correlated with the SID‐MS findings (slope=0.795, intercept=−0.011, r2=0.908) and the performance of the ELISA was satisfactory in terms of recovery (98.5–103.0%) and within‐run (1.4–4.7%) and between‐day (2.8–8.4%) reproducibility. The assay was capable of detecting changes in FIC5.9 during abstinence from drinking in patients with alcohol dependency (p<0.0001).
Conclusions and clinical relevance: The sandwich ELISA developed in this study will be useful for validation of the diagnostic significance of serum FIC5.9 levels in various pathological conditions, including alcoholism.</description><subject>Alcoholism - blood</subject><subject>Biological and medical sciences</subject><subject>Diverse techniques</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Fibrinogen</subject><subject>Fibrinogen alpha C chain</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Liver disease</subject><subject>Male</subject><subject>Mass Spectrometry</subject><subject>Medical research</subject><subject>Middle Aged</subject><subject>Molecular and cellular biology</subject><subject>Peptide Fragments - blood</subject><subject>Proteome - analysis</subject><subject>Proteomics</subject><subject>Reproducibility of Results</subject><subject>Sandwich ELISA</subject><subject>Sensitivity and Specificity</subject><subject>Serum biomarker</subject><subject>Stable isotope-labeled internal standard dilution-MS (SID-MS)</subject><issn>1862-8346</issn><issn>1862-8354</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtv1DAUhSMEoqWwZYksIdRVprbjR7wcpQ8qRgUxQJeW49x03OaFnVDm3-NhhgGxmdW9i--c-zhJ8prgGcGYng3emhnFsceEyifJMckFTfOMs6f7nomj5EUI9xhzRiV-nhxRkgmcU3mcTOfwA5p-aKEbUV8jg4LpqkdnV-hicb2co7r3aFwB4jOVPpwbVLvSu66_gw6ZZlgZVCC7Mq5DtTd3v10qGMGOUKFyjQL4qUWD70foW0CmM806uPAyeVabJsCrXT1Jvl5efCnep4uPV9fFfJFaLrFMc6AKU1wDN4CVUVYQylmtRMZ5yUtJZVZlilFRyspKywSOOp5TJXMwBPPsJDnd-sYNvk8QRt26YKFpTAf9FLQihAghFDlI5kLxXFIqDpNcMSW52kx_-x95308-viBowolkHPNcRWq2pazvQ_BQ68G71vi1JlhvMtabjPU-4yh4s7OdyhaqPf4n1Ai82wEmWNPEYDrrwl-O4XgyZ5FTW-7RNbA-MFZ_-lzM_10i3WpdGOHnXmv8gxYyk1zf3lzp5Yflt-JWZZpmvwDB4sva</recordid><startdate>201104</startdate><enddate>201104</enddate><creator>Noda, Kenta</creator><creator>Sogawa, Kazuyuki</creator><creator>Kikuchi, Wataru</creator><creator>Kiyokawa, Iwao</creator><creator>Miura, Toshihide</creator><creator>Kojima, Ryo</creator><creator>Katayama, Katsuhiro</creator><creator>Kodera, Yoshio</creator><creator>Nomura, Fumio</creator><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</general><general>Wiley</general><general>Wiley Subscription Services, Inc</general><scope>BSCLL</scope><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>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>7QO</scope></search><sort><creationdate>201104</creationdate><title>Development of a sandwich ELISA for the 5.9-kDa fibrinogen alpha C chain fragment detected by serum proteome analysis</title><author>Noda, Kenta ; Sogawa, Kazuyuki ; Kikuchi, Wataru ; Kiyokawa, Iwao ; Miura, Toshihide ; Kojima, Ryo ; Katayama, Katsuhiro ; Kodera, Yoshio ; Nomura, Fumio</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5707-8e29020fe5ae09a9c61254f96355b5b7273d39426b7dc7c460c57582978ea1053</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Alcoholism - blood</topic><topic>Biological and medical sciences</topic><topic>Diverse techniques</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Fibrinogen</topic><topic>Fibrinogen alpha C chain</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Liver disease</topic><topic>Male</topic><topic>Mass Spectrometry</topic><topic>Medical research</topic><topic>Middle Aged</topic><topic>Molecular and cellular biology</topic><topic>Peptide Fragments - blood</topic><topic>Proteome - analysis</topic><topic>Proteomics</topic><topic>Reproducibility of Results</topic><topic>Sandwich ELISA</topic><topic>Sensitivity and Specificity</topic><topic>Serum biomarker</topic><topic>Stable isotope-labeled internal standard dilution-MS (SID-MS)</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Noda, Kenta</creatorcontrib><creatorcontrib>Sogawa, Kazuyuki</creatorcontrib><creatorcontrib>Kikuchi, Wataru</creatorcontrib><creatorcontrib>Kiyokawa, Iwao</creatorcontrib><creatorcontrib>Miura, Toshihide</creatorcontrib><creatorcontrib>Kojima, Ryo</creatorcontrib><creatorcontrib>Katayama, Katsuhiro</creatorcontrib><creatorcontrib>Kodera, Yoshio</creatorcontrib><creatorcontrib>Nomura, Fumio</creatorcontrib><collection>Istex</collection><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>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><jtitle>Proteomics. Clinical applications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Noda, Kenta</au><au>Sogawa, Kazuyuki</au><au>Kikuchi, Wataru</au><au>Kiyokawa, Iwao</au><au>Miura, Toshihide</au><au>Kojima, Ryo</au><au>Katayama, Katsuhiro</au><au>Kodera, Yoshio</au><au>Nomura, Fumio</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a sandwich ELISA for the 5.9-kDa fibrinogen alpha C chain fragment detected by serum proteome analysis</atitle><jtitle>Proteomics. Clinical applications</jtitle><addtitle>Prot. Clin. Appl</addtitle><date>2011-04</date><risdate>2011</risdate><volume>5</volume><issue>3-4</issue><spage>141</spage><epage>146</epage><pages>141-146</pages><issn>1862-8346</issn><eissn>1862-8354</eissn><abstract>Purpose: We previously identified novel biomarker candidates in heavy consumers of alcohol using serum proteome analysis. Among several candidates, a 5.9 kDa peptide identified as a fragment of the fibrinogen alpha C chain (FIC5.9) was the most promising. To move FIC5.9 toward potential diagnostic use, we developed an enzyme immunoassay that enables measurement of serum FIC5.9 levels.
Experimental design: Two monoclonal antibodies specific to the N and C‐termini of the 5.9‐kDa peptide were used to develop a FIC5.9 sandwich ELISA. The assay was evaluated by comparing the results with those obtained by the stable isotope‐labeled dilution mass spectrometry (SID‐MS) using the ClinProt™ system.
Results: The ELISA results correlated with the SID‐MS findings (slope=0.795, intercept=−0.011, r2=0.908) and the performance of the ELISA was satisfactory in terms of recovery (98.5–103.0%) and within‐run (1.4–4.7%) and between‐day (2.8–8.4%) reproducibility. The assay was capable of detecting changes in FIC5.9 during abstinence from drinking in patients with alcohol dependency (p<0.0001).
Conclusions and clinical relevance: The sandwich ELISA developed in this study will be useful for validation of the diagnostic significance of serum FIC5.9 levels in various pathological conditions, including alcoholism.</abstract><cop>Weinheim</cop><pub>WILEY-VCH Verlag</pub><pmid>21360827</pmid><doi>10.1002/prca.201000127</doi><tpages>6</tpages></addata></record> |
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| subjects | Alcoholism - blood Biological and medical sciences Diverse techniques Enzyme-Linked Immunosorbent Assay - methods Fibrinogen Fibrinogen alpha C chain Fundamental and applied biological sciences. Psychology Humans Liver disease Male Mass Spectrometry Medical research Middle Aged Molecular and cellular biology Peptide Fragments - blood Proteome - analysis Proteomics Reproducibility of Results Sandwich ELISA Sensitivity and Specificity Serum biomarker Stable isotope-labeled internal standard dilution-MS (SID-MS) |
| title | Development of a sandwich ELISA for the 5.9-kDa fibrinogen alpha C chain fragment detected by serum proteome analysis |
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