An ELISA for the detection of TIMP-1 based on recombinant single chain Fv fusion proteins
Background: Altered serum levels of TIMP-1 are an indicator of various pathological states. To quantitate TIMP-1 in biological samples, we have previously isolated TIMP-1 specific single-chain Fv fragments (scFvs) using phage-display. In the work presented here, we have used these scFvs to establish...
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| Veröffentlicht in: | Clinica chimica acta 2003-09, Vol.335 (1), p.49-57 |
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| creator | Wozniak, Gordana Obermayr, Eva Jeras, Matjaz Knezevic, Mio Rüker, Florian |
| description | Background: Altered serum levels of TIMP-1 are an indicator of various pathological states. To quantitate TIMP-1 in biological samples, we have previously isolated TIMP-1 specific single-chain Fv fragments (scFvs) using phage-display. In the work presented here, we have used these scFvs to establish a TIMP-1 ELISA based exclusively on recombinant scFv fusion proteins.
Methods: Two distinct TIMP-1 specific scFvs were used as the antigen binding components after being genetically fused to the N-termini of two different fusion protein constructs. One fusion protein, comprising a C
l
domain, serves as a coating reagent, while the second fusion protein with a modified form of bacterial alkaline phosphatase is used as a detection reagent. A double antibody sandwich-ELISA was then established and optimized.
Results: An ELISA for the quantitation of tissue inhibitor of metalloproteinase (TIMP)-1, based entirely on recombinant antibody fragments, was developed as an alternative to assays using polyclonal antisera or monoclonal antibodies. Its performance was shown to compare well with a conventional ELISA. Finally, TIMP-1 concentrations in the sera of sixty healthy individuals were determined.
Conclusion: The assay described here provides a standardized, reliable and readily available means of quantitation of TIMP-1 in a large number of blood samples. |
| doi_str_mv | 10.1016/S0009-8981(03)00281-X |
| format | Article |
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Methods: Two distinct TIMP-1 specific scFvs were used as the antigen binding components after being genetically fused to the N-termini of two different fusion protein constructs. One fusion protein, comprising a C
l
domain, serves as a coating reagent, while the second fusion protein with a modified form of bacterial alkaline phosphatase is used as a detection reagent. A double antibody sandwich-ELISA was then established and optimized.
Results: An ELISA for the quantitation of tissue inhibitor of metalloproteinase (TIMP)-1, based entirely on recombinant antibody fragments, was developed as an alternative to assays using polyclonal antisera or monoclonal antibodies. Its performance was shown to compare well with a conventional ELISA. Finally, TIMP-1 concentrations in the sera of sixty healthy individuals were determined.
Conclusion: The assay described here provides a standardized, reliable and readily available means of quantitation of TIMP-1 in a large number of blood samples.</description><identifier>ISSN: 0009-8981</identifier><identifier>EISSN: 1873-3492</identifier><identifier>DOI: 10.1016/S0009-8981(03)00281-X</identifier><identifier>PMID: 12927684</identifier><identifier>CODEN: CCATAR</identifier><language>eng</language><publisher>Shannon: Elsevier B.V</publisher><subject>Alkaline phosphatase ; Biological and medical sciences ; Cloning, Molecular ; ELISA ; Enzyme-Linked Immunosorbent Assay - methods ; Escherichia coli ; Fusion protein ; Humans ; Immunoglobulin Fragments - immunology ; Immunoglobulin Variable Region - immunology ; Investigative techniques, diagnostic techniques (general aspects) ; Medical sciences ; Miscellaneous. Technology ; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques ; Phage display ; Recombinant Fusion Proteins - biosynthesis ; Single-chain Fv ; TIMP-1 ; Tissue Inhibitor of Metalloproteinase-1 - analysis ; Tissue Inhibitor of Metalloproteinase-1 - blood ; Tissue Inhibitor of Metalloproteinase-1 - immunology</subject><ispartof>Clinica chimica acta, 2003-09, Vol.335 (1), p.49-57</ispartof><rights>2003 Elsevier B.V.</rights><rights>2004 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c391t-afffe3b93f709938859435231cf14a506c27a5d8aa41bc1feab2c2eebd1a5ca63</citedby><cites>FETCH-LOGICAL-c391t-afffe3b93f709938859435231cf14a506c27a5d8aa41bc1feab2c2eebd1a5ca63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S000989810300281X$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15080308$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12927684$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wozniak, Gordana</creatorcontrib><creatorcontrib>Obermayr, Eva</creatorcontrib><creatorcontrib>Jeras, Matjaz</creatorcontrib><creatorcontrib>Knezevic, Mio</creatorcontrib><creatorcontrib>Rüker, Florian</creatorcontrib><title>An ELISA for the detection of TIMP-1 based on recombinant single chain Fv fusion proteins</title><title>Clinica chimica acta</title><addtitle>Clin Chim Acta</addtitle><description>Background: Altered serum levels of TIMP-1 are an indicator of various pathological states. To quantitate TIMP-1 in biological samples, we have previously isolated TIMP-1 specific single-chain Fv fragments (scFvs) using phage-display. In the work presented here, we have used these scFvs to establish a TIMP-1 ELISA based exclusively on recombinant scFv fusion proteins.
Methods: Two distinct TIMP-1 specific scFvs were used as the antigen binding components after being genetically fused to the N-termini of two different fusion protein constructs. One fusion protein, comprising a C
l
domain, serves as a coating reagent, while the second fusion protein with a modified form of bacterial alkaline phosphatase is used as a detection reagent. A double antibody sandwich-ELISA was then established and optimized.
Results: An ELISA for the quantitation of tissue inhibitor of metalloproteinase (TIMP)-1, based entirely on recombinant antibody fragments, was developed as an alternative to assays using polyclonal antisera or monoclonal antibodies. Its performance was shown to compare well with a conventional ELISA. Finally, TIMP-1 concentrations in the sera of sixty healthy individuals were determined.
Conclusion: The assay described here provides a standardized, reliable and readily available means of quantitation of TIMP-1 in a large number of blood samples.</description><subject>Alkaline phosphatase</subject><subject>Biological and medical sciences</subject><subject>Cloning, Molecular</subject><subject>ELISA</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Escherichia coli</subject><subject>Fusion protein</subject><subject>Humans</subject><subject>Immunoglobulin Fragments - immunology</subject><subject>Immunoglobulin Variable Region - immunology</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Medical sciences</subject><subject>Miscellaneous. Technology</subject><subject>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</subject><subject>Phage display</subject><subject>Recombinant Fusion Proteins - biosynthesis</subject><subject>Single-chain Fv</subject><subject>TIMP-1</subject><subject>Tissue Inhibitor of Metalloproteinase-1 - analysis</subject><subject>Tissue Inhibitor of Metalloproteinase-1 - blood</subject><subject>Tissue Inhibitor of Metalloproteinase-1 - immunology</subject><issn>0009-8981</issn><issn>1873-3492</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1PGzEQhq2qiITAT2jlSxEcttjr_bBPVYSgRAoqEkFKT9asd9y42nipvUHi39chETlyGs3oeWdGDyFfOPvOGa-uHhljKpNK8gsmLhnLJc-Wn8iYy1pkolD5ZzJ-R0bkJMa_qS1YxY_JiOcqrytZjMnvqac389njlNo-0GGFtMUBzeB6T3tLF7P7h4zTBiK2NI0Cmn7dOA9-oNH5Px1SswLn6e0LtZu4TT2HfkDn4yk5stBFPNvXCXm6vVlc32XzXz9n19N5ZoTiQwbWWhSNErZmSgkpS1WIMhfcWF5AySqT11C2EqDgjeEWoclNjti0HEoDlZiQ893edPjfBuOg1y4a7Drw2G-irkVZS1EUCSx3oAl9jAGtfg5uDeFVc6a3TvWbU70VppnQb071MuW-7g9smjW2h9ReYgK-7QGIBjobwBsXD1zJJBNMJu7HjsOk48Vh0NE49AZbl7wOuu3dB6_8B0lqkrc</recordid><startdate>20030901</startdate><enddate>20030901</enddate><creator>Wozniak, Gordana</creator><creator>Obermayr, Eva</creator><creator>Jeras, Matjaz</creator><creator>Knezevic, Mio</creator><creator>Rüker, Florian</creator><general>Elsevier B.V</general><general>Elsevier</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>7X8</scope></search><sort><creationdate>20030901</creationdate><title>An ELISA for the detection of TIMP-1 based on recombinant single chain Fv fusion proteins</title><author>Wozniak, Gordana ; Obermayr, Eva ; Jeras, Matjaz ; Knezevic, Mio ; Rüker, Florian</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c391t-afffe3b93f709938859435231cf14a506c27a5d8aa41bc1feab2c2eebd1a5ca63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Alkaline phosphatase</topic><topic>Biological and medical sciences</topic><topic>Cloning, Molecular</topic><topic>ELISA</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Escherichia coli</topic><topic>Fusion protein</topic><topic>Humans</topic><topic>Immunoglobulin Fragments - immunology</topic><topic>Immunoglobulin Variable Region - immunology</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Medical sciences</topic><topic>Miscellaneous. Technology</topic><topic>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</topic><topic>Phage display</topic><topic>Recombinant Fusion Proteins - biosynthesis</topic><topic>Single-chain Fv</topic><topic>TIMP-1</topic><topic>Tissue Inhibitor of Metalloproteinase-1 - analysis</topic><topic>Tissue Inhibitor of Metalloproteinase-1 - blood</topic><topic>Tissue Inhibitor of Metalloproteinase-1 - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wozniak, Gordana</creatorcontrib><creatorcontrib>Obermayr, Eva</creatorcontrib><creatorcontrib>Jeras, Matjaz</creatorcontrib><creatorcontrib>Knezevic, Mio</creatorcontrib><creatorcontrib>Rüker, Florian</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>MEDLINE - Academic</collection><jtitle>Clinica chimica acta</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wozniak, Gordana</au><au>Obermayr, Eva</au><au>Jeras, Matjaz</au><au>Knezevic, Mio</au><au>Rüker, Florian</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An ELISA for the detection of TIMP-1 based on recombinant single chain Fv fusion proteins</atitle><jtitle>Clinica chimica acta</jtitle><addtitle>Clin Chim Acta</addtitle><date>2003-09-01</date><risdate>2003</risdate><volume>335</volume><issue>1</issue><spage>49</spage><epage>57</epage><pages>49-57</pages><issn>0009-8981</issn><eissn>1873-3492</eissn><coden>CCATAR</coden><abstract>Background: Altered serum levels of TIMP-1 are an indicator of various pathological states. To quantitate TIMP-1 in biological samples, we have previously isolated TIMP-1 specific single-chain Fv fragments (scFvs) using phage-display. In the work presented here, we have used these scFvs to establish a TIMP-1 ELISA based exclusively on recombinant scFv fusion proteins.
Methods: Two distinct TIMP-1 specific scFvs were used as the antigen binding components after being genetically fused to the N-termini of two different fusion protein constructs. One fusion protein, comprising a C
l
domain, serves as a coating reagent, while the second fusion protein with a modified form of bacterial alkaline phosphatase is used as a detection reagent. A double antibody sandwich-ELISA was then established and optimized.
Results: An ELISA for the quantitation of tissue inhibitor of metalloproteinase (TIMP)-1, based entirely on recombinant antibody fragments, was developed as an alternative to assays using polyclonal antisera or monoclonal antibodies. Its performance was shown to compare well with a conventional ELISA. Finally, TIMP-1 concentrations in the sera of sixty healthy individuals were determined.
Conclusion: The assay described here provides a standardized, reliable and readily available means of quantitation of TIMP-1 in a large number of blood samples.</abstract><cop>Shannon</cop><pub>Elsevier B.V</pub><pmid>12927684</pmid><doi>10.1016/S0009-8981(03)00281-X</doi><tpages>9</tpages></addata></record> |
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| subjects | Alkaline phosphatase Biological and medical sciences Cloning, Molecular ELISA Enzyme-Linked Immunosorbent Assay - methods Escherichia coli Fusion protein Humans Immunoglobulin Fragments - immunology Immunoglobulin Variable Region - immunology Investigative techniques, diagnostic techniques (general aspects) Medical sciences Miscellaneous. Technology Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Phage display Recombinant Fusion Proteins - biosynthesis Single-chain Fv TIMP-1 Tissue Inhibitor of Metalloproteinase-1 - analysis Tissue Inhibitor of Metalloproteinase-1 - blood Tissue Inhibitor of Metalloproteinase-1 - immunology |
| title | An ELISA for the detection of TIMP-1 based on recombinant single chain Fv fusion proteins |
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