Synthesis of a cortisol-biotin conjugate and evaluation as a tracer in an immunoassay for salivary cortisol measurement
Identifieur interne : 001460 ( Istex/Corpus ); précédent : 001459; suivant : 001461Synthesis of a cortisol-biotin conjugate and evaluation as a tracer in an immunoassay for salivary cortisol measurement
Auteurs : R. A. Dressendörfer ; C. Kirschbaum ; W. Rohde ; F. Stahl ; C. J. StrasburgerSource :
- Journal of Steroid Biochemistry and Molecular Biology [ 0960-0760 ] ; 1992.
Abstract
Cortisol 3-(o-carboxymethyl)oxime (C3-CMO) and a commercially available biotin-hydrazide derivative were used to synthesize a C3-CMO-biotin conjugate. C3-CMO was converted into a N-hydroxysuccinimide ester derivative which in a second reaction step was allowed to interact with the hydrazide derivative of biotin. This simple-to-perform synthesis yielded a conjugate suitable for use as a tracer in immunoassays for cortisol measurement. Employing biotin as the primary probe in a competitive solid phase immunoassay allows for variable end point determination by means of commercially available labeled avidin or streptavidin derivatives. Streptavidin-Europium was used in conjunction with the DELFIA-system for time-resolved fluorometric end point measurement (TR-FIA) throughout the study. In addition, colorimetric end point determination (ELISA) using streptavidin-alkaline phosphatase as a secondary probe was established and evaluated. Both forms of this non-isotopic assay showed excellent correlation with a commercially available radioimmunoassay adapted for salivary cortisol measurement. The lower detection limit was 0.43 nM for a 50 μl salivary sample. The intra-assay coefficient of variation was 6.7, 4.7 and 4.0% at cortisol concentrations of 2.2, 5.5 and 13.2 nM, respectively (n = 37), and the corresponding inter-assay coefficients of variation were 9.0, 8.6 and 7.1% (n = 50). The competitive immunoassay requires 1.5 h incubation time and shows robust and reproducible performance. The C3-CMO-biotin conjugate allows for sensitive and flexible end point determination of salivary cortisol levels in immunoassays.
Url:
DOI: 10.1016/0960-0760(92)90294-S
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A." last="Dressendörfer">R. A. Dressendörfer</name><affiliation><mods:affiliation>Medizinische Klinik, Klinikum Innenstadt der Ludwig-Maximilians-Universität, Ziemssenstraβe 1, 8000 München 2, Germany</mods:affiliation></affiliation></author><author><name sortKey="Kirschbaum, C" sort="Kirschbaum, C" uniqKey="Kirschbaum C" first="C." last="Kirschbaum">C. Kirschbaum</name><affiliation><mods:affiliation>Fachbereich I Psychologie, Universität Trier, 5500 Trier-Tarforst, Germany</mods:affiliation></affiliation></author><author><name sortKey="Rohde, W" sort="Rohde, W" uniqKey="Rohde W" first="W." last="Rohde">W. Rohde</name><affiliation><mods:affiliation>Institut für Experimentelle Endokrinologie, Bereich Medizin (Charité) der Humboldt-Universität zu Berlin, Schumannstraβe 20/21, 1040 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Stahl, F" sort="Stahl, F" uniqKey="Stahl F" first="F." last="Stahl">F. Stahl</name><affiliation><mods:affiliation>Institut für Experimentelle Endokrinologie, Bereich Medizin (Charité) der Humboldt-Universität zu Berlin, Schumannstraβe 20/21, 1040 Berlin, Germany</mods:affiliation></affiliation></author><author><name sortKey="Strasburger, C J" sort="Strasburger, C J" uniqKey="Strasburger C" first="C. J." last="Strasburger">C. J. Strasburger</name><affiliation><mods:affiliation>To whom correspondence should be addressed.</mods:affiliation></affiliation><affiliation><mods:affiliation>Medizinische Klinik, Klinikum Innenstadt der Ludwig-Maximilians-Universität, Ziemssenstraβe 1, 8000 München 2, Germany</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Steroid Biochemistry and Molecular Biology</title><title level="j" type="abbrev">SBMB</title><idno type="ISSN">0960-0760</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1992">1992</date><biblScope unit="volume">43</biblScope><biblScope unit="issue">7</biblScope><biblScope unit="page" from="683">683</biblScope><biblScope unit="page" to="692">692</biblScope></imprint><idno type="ISSN">0960-0760</idno></series><idno type="istex">73592A6264FB200B44EB3A3DC6BBCFB83BF3C8B7</idno><idno type="DOI">10.1016/0960-0760(92)90294-S</idno><idno type="PII">0960-0760(92)90294-S</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0960-0760</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Cortisol 3-(o-carboxymethyl)oxime (C3-CMO) and a commercially available biotin-hydrazide derivative were used to synthesize a C3-CMO-biotin conjugate. C3-CMO was converted into a N-hydroxysuccinimide ester derivative which in a second reaction step was allowed to interact with the hydrazide derivative of biotin. This simple-to-perform synthesis yielded a conjugate suitable for use as a tracer in immunoassays for cortisol measurement. Employing biotin as the primary probe in a competitive solid phase immunoassay allows for variable end point determination by means of commercially available labeled avidin or streptavidin derivatives. Streptavidin-Europium was used in conjunction with the DELFIA-system for time-resolved fluorometric end point measurement (TR-FIA) throughout the study. In addition, colorimetric end point determination (ELISA) using streptavidin-alkaline phosphatase as a secondary probe was established and evaluated. Both forms of this non-isotopic assay showed excellent correlation with a commercially available radioimmunoassay adapted for salivary cortisol measurement. The lower detection limit was 0.43 nM for a 50 μl salivary sample. The intra-assay coefficient of variation was 6.7, 4.7 and 4.0% at cortisol concentrations of 2.2, 5.5 and 13.2 nM, respectively (n = 37), and the corresponding inter-assay coefficients of variation were 9.0, 8.6 and 7.1% (n = 50). The competitive immunoassay requires 1.5 h incubation time and shows robust and reproducible performance. The C3-CMO-biotin conjugate allows for sensitive and flexible end point determination of salivary cortisol levels in immunoassays.</div></front></TEI><istex><corpusName>elsevier</corpusName><author><json:item><name>R.A. Dressendörfer</name><affiliations><json:string>Medizinische Klinik, Klinikum Innenstadt der Ludwig-Maximilians-Universität, Ziemssenstraβe 1, 8000 München 2, Germany</json:string></affiliations></json:item><json:item><name>C. Kirschbaum</name><affiliations><json:string>Fachbereich I Psychologie, Universität Trier, 5500 Trier-Tarforst, Germany</json:string></affiliations></json:item><json:item><name>W. Rohde</name><affiliations><json:string>Institut für Experimentelle Endokrinologie, Bereich Medizin (Charité) der Humboldt-Universität zu Berlin, Schumannstraβe 20/21, 1040 Berlin, Germany</json:string></affiliations></json:item><json:item><name>F. Stahl</name><affiliations><json:string>Institut für Experimentelle Endokrinologie, Bereich Medizin (Charité) der Humboldt-Universität zu Berlin, Schumannstraβe 20/21, 1040 Berlin, Germany</json:string></affiliations></json:item><json:item><name>C.J. Strasburger</name><affiliations><json:string>To whom correspondence should be addressed.</json:string><json:string>Medizinische Klinik, Klinikum Innenstadt der Ludwig-Maximilians-Universität, Ziemssenstraβe 1, 8000 München 2, Germany</json:string></affiliations></json:item></author><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>Cortisol 3-(o-carboxymethyl)oxime (C3-CMO) and a commercially available biotin-hydrazide derivative were used to synthesize a C3-CMO-biotin conjugate. C3-CMO was converted into a N-hydroxysuccinimide ester derivative which in a second reaction step was allowed to interact with the hydrazide derivative of biotin. This simple-to-perform synthesis yielded a conjugate suitable for use as a tracer in immunoassays for cortisol measurement. Employing biotin as the primary probe in a competitive solid phase immunoassay allows for variable end point determination by means of commercially available labeled avidin or streptavidin derivatives. Streptavidin-Europium was used in conjunction with the DELFIA-system for time-resolved fluorometric end point measurement (TR-FIA) throughout the study. In addition, colorimetric end point determination (ELISA) using streptavidin-alkaline phosphatase as a secondary probe was established and evaluated. Both forms of this non-isotopic assay showed excellent correlation with a commercially available radioimmunoassay adapted for salivary cortisol measurement. The lower detection limit was 0.43 nM for a 50 μl salivary sample. The intra-assay coefficient of variation was 6.7, 4.7 and 4.0% at cortisol concentrations of 2.2, 5.5 and 13.2 nM, respectively (n = 37), and the corresponding inter-assay coefficients of variation were 9.0, 8.6 and 7.1% (n = 50). The competitive immunoassay requires 1.5 h incubation time and shows robust and reproducible performance. The C3-CMO-biotin conjugate allows for sensitive and flexible end point determination of salivary cortisol levels in immunoassays.</abstract><qualityIndicators><score>6.713</score><pdfVersion>1.2</pdfVersion><pdfPageSize>533 x 771 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>0</keywordCount><abstractCharCount>1634</abstractCharCount><pdfWordCount>4049</pdfWordCount><pdfCharCount>27041</pdfCharCount><pdfPageCount>10</pdfPageCount><abstractWordCount>222</abstractWordCount></qualityIndicators><title>Synthesis of a cortisol-biotin conjugate and evaluation as a tracer in an immunoassay for salivary cortisol measurement</title><pii><json:string>0960-0760(92)90294-S</json:string></pii><refBibs><json:item><author><json:item><name>B.P. Murphy</name></json:item><json:item><name>W. Engelberg</name></json:item><json:item><name>C.J. 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C3-CMO was converted into a N-hydroxysuccinimide ester derivative which in a second reaction step was allowed to interact with the hydrazide derivative of biotin. This simple-to-perform synthesis yielded a conjugate suitable for use as a tracer in immunoassays for cortisol measurement. Employing biotin as the primary probe in a competitive solid phase immunoassay allows for variable end point determination by means of commercially available labeled avidin or streptavidin derivatives. Streptavidin-Europium was used in conjunction with the DELFIA-system for time-resolved fluorometric end point measurement (TR-FIA) throughout the study. In addition, colorimetric end point determination (ELISA) using streptavidin-alkaline phosphatase as a secondary probe was established and evaluated. Both forms of this non-isotopic assay showed excellent correlation with a commercially available radioimmunoassay adapted for salivary cortisol measurement. The lower detection limit was 0.43 nM for a 50 μl salivary sample. The intra-assay coefficient of variation was 6.7, 4.7 and 4.0% at cortisol concentrations of 2.2, 5.5 and 13.2 nM, respectively (n = 37), and the corresponding inter-assay coefficients of variation were 9.0, 8.6 and 7.1% (n = 50). The competitive immunoassay requires 1.5 h incubation time and shows robust and reproducible performance. The C3-CMO-biotin conjugate allows for sensitive and flexible end point determination of salivary cortisol levels in immunoassays.</p></abstract></profileDesc><revisionDesc><change when="1992">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/document/73592A6264FB200B44EB3A3DC6BBCFB83BF3C8B7/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>SBMB</jid><aid>9290294S</aid><ce:pii>0960-0760(92)90294-S</ce:pii><ce:doi>10.1016/0960-0760(92)90294-S</ce:doi><ce:copyright type="unknown" year="1992"></ce:copyright></item-info><head><ce:title>Synthesis of a cortisol-biotin conjugate and evaluation as a tracer in an immunoassay for salivary cortisol measurement</ce:title><ce:author-group><ce:author><ce:given-name>R.A.</ce:given-name><ce:surname>Dressendörfer</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>1</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>C.</ce:given-name><ce:surname>Kirschbaum</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>2</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>W.</ce:given-name><ce:surname>Rohde</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>3</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>F.</ce:given-name><ce:surname>Stahl</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>3</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>C.J.</ce:given-name><ce:surname>Strasburger</ce:surname><ce:cross-ref refid="COR1"><ce:sup>∗</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF1"><ce:sup>1</ce:sup></ce:cross-ref></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Medizinische Klinik, Klinikum Innenstadt der Ludwig-Maximilians-Universität, Ziemssenstraβe 1, 8000 München 2, Germany</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>Fachbereich I Psychologie, Universität Trier, 5500 Trier-Tarforst, Germany</ce:textfn></ce:affiliation><ce:affiliation id="AFF3"><ce:label>c</ce:label><ce:textfn>Institut für Experimentelle Endokrinologie, Bereich Medizin (Charité) der Humboldt-Universität zu Berlin, Schumannstraβe 20/21, 1040 Berlin, Germany</ce:textfn></ce:affiliation><ce:correspondence id="COR1"><ce:label>∗</ce:label><ce:text>To whom correspondence should be addressed.</ce:text></ce:correspondence></ce:author-group><ce:date-received day="2" month="12" year="1991"></ce:date-received><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>Cortisol 3-(<ce:italic>o</ce:italic>-carboxymethyl)oxime (C3-CMO) and a commercially available biotin-hydrazide derivative were used to synthesize a C3-CMO-biotin conjugate. C3-CMO was converted into a <ce:italic>N</ce:italic>-hydroxysuccinimide ester derivative which in a second reaction step was allowed to interact with the hydrazide derivative of biotin. This simple-to-perform synthesis yielded a conjugate suitable for use as a tracer in immunoassays for cortisol measurement. Employing biotin as the primary probe in a competitive solid phase immunoassay allows for variable end point determination by means of commercially available labeled avidin or streptavidin derivatives. Streptavidin-Europium was used in conjunction with the DELFIA-system for time-resolved fluorometric end point measurement (TR-FIA) throughout the study. In addition, colorimetric end point determination (ELISA) using streptavidin-alkaline phosphatase as a secondary probe was established and evaluated. Both forms of this non-isotopic assay showed excellent correlation with a commercially available radioimmunoassay adapted for salivary cortisol measurement. The lower detection limit was 0.43 nM for a 50 μl salivary sample. The intra-assay coefficient of variation was 6.7, 4.7 and 4.0% at cortisol concentrations of 2.2, 5.5 and 13.2 nM, respectively (<ce:italic>n</ce:italic> = 37), and the corresponding inter-assay coefficients of variation were 9.0, 8.6 and 7.1% (<ce:italic>n</ce:italic> = 50). The competitive immunoassay requires 1.5 h incubation time and shows robust and reproducible performance. The C3-CMO-biotin conjugate allows for sensitive and flexible end point determination of salivary cortisol levels in immunoassays.</ce:simple-para></ce:abstract-sec></ce:abstract></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Synthesis of a cortisol-biotin conjugate and evaluation as a tracer in an immunoassay for salivary cortisol measurement</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Synthesis of a cortisol-biotin conjugate and evaluation as a tracer in an immunoassay for salivary cortisol measurement</title></titleInfo><name type="personal"><namePart type="given">R.A.</namePart><namePart type="family">Dressendörfer</namePart><affiliation>Medizinische Klinik, Klinikum Innenstadt der Ludwig-Maximilians-Universität, Ziemssenstraβe 1, 8000 München 2, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">C.</namePart><namePart type="family">Kirschbaum</namePart><affiliation>Fachbereich I Psychologie, Universität Trier, 5500 Trier-Tarforst, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">W.</namePart><namePart type="family">Rohde</namePart><affiliation>Institut für Experimentelle Endokrinologie, Bereich Medizin (Charité) der Humboldt-Universität zu Berlin, Schumannstraβe 20/21, 1040 Berlin, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">F.</namePart><namePart type="family">Stahl</namePart><affiliation>Institut für Experimentelle Endokrinologie, Bereich Medizin (Charité) der Humboldt-Universität zu Berlin, Schumannstraβe 20/21, 1040 Berlin, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">C.J.</namePart><namePart type="family">Strasburger</namePart><affiliation>To whom correspondence should be addressed.</affiliation><affiliation>Medizinische Klinik, Klinikum Innenstadt der Ludwig-Maximilians-Universität, Ziemssenstraβe 1, 8000 München 2, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article"></genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1992</dateIssued><copyrightDate encoding="w3cdtf">1992</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">Cortisol 3-(o-carboxymethyl)oxime (C3-CMO) and a commercially available biotin-hydrazide derivative were used to synthesize a C3-CMO-biotin conjugate. C3-CMO was converted into a N-hydroxysuccinimide ester derivative which in a second reaction step was allowed to interact with the hydrazide derivative of biotin. This simple-to-perform synthesis yielded a conjugate suitable for use as a tracer in immunoassays for cortisol measurement. Employing biotin as the primary probe in a competitive solid phase immunoassay allows for variable end point determination by means of commercially available labeled avidin or streptavidin derivatives. Streptavidin-Europium was used in conjunction with the DELFIA-system for time-resolved fluorometric end point measurement (TR-FIA) throughout the study. In addition, colorimetric end point determination (ELISA) using streptavidin-alkaline phosphatase as a secondary probe was established and evaluated. Both forms of this non-isotopic assay showed excellent correlation with a commercially available radioimmunoassay adapted for salivary cortisol measurement. The lower detection limit was 0.43 nM for a 50 μl salivary sample. The intra-assay coefficient of variation was 6.7, 4.7 and 4.0% at cortisol concentrations of 2.2, 5.5 and 13.2 nM, respectively (n = 37), and the corresponding inter-assay coefficients of variation were 9.0, 8.6 and 7.1% (n = 50). The competitive immunoassay requires 1.5 h incubation time and shows robust and reproducible performance. The C3-CMO-biotin conjugate allows for sensitive and flexible end point determination of salivary cortisol levels in immunoassays.</abstract><relatedItem type="host"><titleInfo><title>Journal of Steroid Biochemistry and Molecular Biology</title></titleInfo><titleInfo type="abbreviated"><title>SBMB</title></titleInfo><genre type="journal">journal</genre><originInfo><dateIssued encoding="w3cdtf">199212</dateIssued></originInfo><identifier type="ISSN">0960-0760</identifier><identifier type="PII">S0960-0760(00)X0128-8</identifier><part><date>199212</date><detail type="volume"><number>43</number><caption>vol.</caption></detail><detail type="issue"><number>7</number><caption>no.</caption></detail><extent unit="issue pages"><start>599</start><end>743</end></extent><extent unit="pages"><start>683</start><end>692</end></extent></part></relatedItem><identifier type="istex">73592A6264FB200B44EB3A3DC6BBCFB83BF3C8B7</identifier><identifier type="DOI">10.1016/0960-0760(92)90294-S</identifier><identifier type="PII">0960-0760(92)90294-S</identifier><recordInfo><recordContentSource>ELSEVIER</recordContentSource></recordInfo></mods></metadata></istex></record>Pour manipuler ce document sous Unix (Dilib)
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