Europium-labelled streptavidin as a highly sensitive universal label: Indirect time-resolved immunofluorometry of FSH and TSH
Labelling of streptavidin with a fluorogenic europium ion was optimized with the aim of obtaining a universal, stable and highly sensitive non-isotopic label for time-resolved fluorometric immunoassays (TR-FIA) based on dissociative fluorescence enhancement (DELFIA). Even the conjugation of all the...
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| Veröffentlicht in: | Journal of immunological methods 1992-05, Vol.149 (2), p.247-253 |
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| creator | Suonpää, M. Markela, E. Ståhlberg, T. Hemmilä, I. |
| description | Labelling of streptavidin with a fluorogenic europium ion was optimized with the aim of obtaining a universal, stable and highly sensitive non-isotopic label for time-resolved fluorometric immunoassays (TR-FIA) based on dissociative fluorescence enhancement (DELFIA). Even the conjugation of all the free amino groups of streptavidin with Eu chelates had only a minor effect on the binding capacity of the protein or its affinity. The labelled streptavidin was evaluated in indirect time-resolved immunofluorometric assays of human follicle and thyroid stimulating hormones (FSH and TSH). The interassay imprecision was below 3% within the concentration range from 2.5 to 94 U/I for the FSH samples and below 5% in the range from 2.4 to 35 mIU/l for the TSH samples. The detection limits of the assays for FSH and TSH were 0.05–0.10 U/l and 0.01–0.025 mIU/l, respectively, when a CV of 15% was regarded as the acceptable upper limit of imprecision. The results obtained by the indirect assays correlated closely with those obtained by corresponding direct sandwich assays. The model assays demonstrated the utility of Eu-labelled streptavidin as a universal reagent for immunoassays requiring a wide dynamic range and high sensitivity. |
| doi_str_mv | 10.1016/0022-1759(92)90256-S |
| format | Article |
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Even the conjugation of all the free amino groups of streptavidin with Eu chelates had only a minor effect on the binding capacity of the protein or its affinity. The labelled streptavidin was evaluated in indirect time-resolved immunofluorometric assays of human follicle and thyroid stimulating hormones (FSH and TSH). The interassay imprecision was below 3% within the concentration range from 2.5 to 94 U/I for the FSH samples and below 5% in the range from 2.4 to 35 mIU/l for the TSH samples. The detection limits of the assays for FSH and TSH were 0.05–0.10 U/l and 0.01–0.025 mIU/l, respectively, when a CV of 15% was regarded as the acceptable upper limit of imprecision. The results obtained by the indirect assays correlated closely with those obtained by corresponding direct sandwich assays. The model assays demonstrated the utility of Eu-labelled streptavidin as a universal reagent for immunoassays requiring a wide dynamic range and high sensitivity.</description><identifier>ISSN: 0022-1759</identifier><identifier>EISSN: 1872-7905</identifier><identifier>DOI: 10.1016/0022-1759(92)90256-S</identifier><identifier>PMID: 1593136</identifier><identifier>CODEN: JIMMBG</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Bacterial Proteins ; Biological and medical sciences ; DELFIA ; Europium ; Fluoroimmunoassay ; Fluoroimmunoassay - methods ; Follicle stimulating hormone ; Follicle Stimulating Hormone - analysis ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Humans ; Isotopes ; Molecular immunology ; Sensitivity and Specificity ; Streptavidin ; Techniques ; Thyroid stimulating hormone ; Thyrotropin - analysis ; Time Factors ; Time-resolved fluorometry</subject><ispartof>Journal of immunological methods, 1992-05, Vol.149 (2), p.247-253</ispartof><rights>1992</rights><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0022-1759(92)90256-S$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27923,27924,45994</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5297551$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1593136$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Suonpää, M.</creatorcontrib><creatorcontrib>Markela, E.</creatorcontrib><creatorcontrib>Ståhlberg, T.</creatorcontrib><creatorcontrib>Hemmilä, I.</creatorcontrib><title>Europium-labelled streptavidin as a highly sensitive universal label: Indirect time-resolved immunofluorometry of FSH and TSH</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>Labelling of streptavidin with a fluorogenic europium ion was optimized with the aim of obtaining a universal, stable and highly sensitive non-isotopic label for time-resolved fluorometric immunoassays (TR-FIA) based on dissociative fluorescence enhancement (DELFIA). Even the conjugation of all the free amino groups of streptavidin with Eu chelates had only a minor effect on the binding capacity of the protein or its affinity. The labelled streptavidin was evaluated in indirect time-resolved immunofluorometric assays of human follicle and thyroid stimulating hormones (FSH and TSH). The interassay imprecision was below 3% within the concentration range from 2.5 to 94 U/I for the FSH samples and below 5% in the range from 2.4 to 35 mIU/l for the TSH samples. The detection limits of the assays for FSH and TSH were 0.05–0.10 U/l and 0.01–0.025 mIU/l, respectively, when a CV of 15% was regarded as the acceptable upper limit of imprecision. The results obtained by the indirect assays correlated closely with those obtained by corresponding direct sandwich assays. The model assays demonstrated the utility of Eu-labelled streptavidin as a universal reagent for immunoassays requiring a wide dynamic range and high sensitivity.</description><subject>Bacterial Proteins</subject><subject>Biological and medical sciences</subject><subject>DELFIA</subject><subject>Europium</subject><subject>Fluoroimmunoassay</subject><subject>Fluoroimmunoassay - methods</subject><subject>Follicle stimulating hormone</subject><subject>Follicle Stimulating Hormone - analysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Humans</subject><subject>Isotopes</subject><subject>Molecular immunology</subject><subject>Sensitivity and Specificity</subject><subject>Streptavidin</subject><subject>Techniques</subject><subject>Thyroid stimulating hormone</subject><subject>Thyrotropin - analysis</subject><subject>Time Factors</subject><subject>Time-resolved fluorometry</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1rGzEQhkVoSJ00_6AFHUpJD5vqY_WVQ6GEpA4EenB6FvJqtlHRal1p1-BD_3vl2OTaywzM-8zAOy9C7ym5poTKL4Qw1lAlzJVhnw1hQjarE7SgWrFGGSLeoMUr8hadl_KbEEKJJGfojArDKZcL9PduzuMmzEMT3RpiBI_LlGEzuW3wIWFXsMPP4ddz3OECqYQpbAHPqdZcXMQvWzf4IfmQoZvwFAZoMpQxbuupMAxzGvs4j3kcYMo7PPb4frXELnn8tFq-Q6e9iwUuj_0C_by_e7pdNo8_vj_cfntsgDM6Na0UTGnoneN6rc26Z9D2SvG25dp5CUJUz7pvDRdAW0WNrGNgXkvwvAXOL9Cnw91NHv_MUCY7hNJVuy7BOBermJGUa_1fkErOiDKygh-O4LwewNtNDoPLO3t8bNU_HnVXOhf77FIXyismmFFC0Ip9PWBQ3W8DZFu6AKmDwzutH4OlxO7ztvsw7T5Ma5h9yduu-D8tYpz6</recordid><startdate>19920518</startdate><enddate>19920518</enddate><creator>Suonpää, M.</creator><creator>Markela, E.</creator><creator>Ståhlberg, T.</creator><creator>Hemmilä, I.</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>7T5</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>19920518</creationdate><title>Europium-labelled streptavidin as a highly sensitive universal label: Indirect time-resolved immunofluorometry of FSH and TSH</title><author>Suonpää, M. ; Markela, E. ; Ståhlberg, T. ; Hemmilä, I.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-e321t-465278efaa38b89bf2e4f7734438ad6e558728f4935e1471968ade2d86ed34e33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Bacterial Proteins</topic><topic>Biological and medical sciences</topic><topic>DELFIA</topic><topic>Europium</topic><topic>Fluoroimmunoassay</topic><topic>Fluoroimmunoassay - methods</topic><topic>Follicle stimulating hormone</topic><topic>Follicle Stimulating Hormone - analysis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Humans</topic><topic>Isotopes</topic><topic>Molecular immunology</topic><topic>Sensitivity and Specificity</topic><topic>Streptavidin</topic><topic>Techniques</topic><topic>Thyroid stimulating hormone</topic><topic>Thyrotropin - analysis</topic><topic>Time Factors</topic><topic>Time-resolved fluorometry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Suonpää, M.</creatorcontrib><creatorcontrib>Markela, E.</creatorcontrib><creatorcontrib>Ståhlberg, T.</creatorcontrib><creatorcontrib>Hemmilä, I.</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>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Suonpää, M.</au><au>Markela, E.</au><au>Ståhlberg, T.</au><au>Hemmilä, I.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Europium-labelled streptavidin as a highly sensitive universal label: Indirect time-resolved immunofluorometry of FSH and TSH</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>1992-05-18</date><risdate>1992</risdate><volume>149</volume><issue>2</issue><spage>247</spage><epage>253</epage><pages>247-253</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>Labelling of streptavidin with a fluorogenic europium ion was optimized with the aim of obtaining a universal, stable and highly sensitive non-isotopic label for time-resolved fluorometric immunoassays (TR-FIA) based on dissociative fluorescence enhancement (DELFIA). Even the conjugation of all the free amino groups of streptavidin with Eu chelates had only a minor effect on the binding capacity of the protein or its affinity. The labelled streptavidin was evaluated in indirect time-resolved immunofluorometric assays of human follicle and thyroid stimulating hormones (FSH and TSH). The interassay imprecision was below 3% within the concentration range from 2.5 to 94 U/I for the FSH samples and below 5% in the range from 2.4 to 35 mIU/l for the TSH samples. The detection limits of the assays for FSH and TSH were 0.05–0.10 U/l and 0.01–0.025 mIU/l, respectively, when a CV of 15% was regarded as the acceptable upper limit of imprecision. The results obtained by the indirect assays correlated closely with those obtained by corresponding direct sandwich assays. The model assays demonstrated the utility of Eu-labelled streptavidin as a universal reagent for immunoassays requiring a wide dynamic range and high sensitivity.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>1593136</pmid><doi>10.1016/0022-1759(92)90256-S</doi><tpages>7</tpages></addata></record> |
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| ispartof | Journal of immunological methods, 1992-05, Vol.149 (2), p.247-253 |
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| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | Bacterial Proteins Biological and medical sciences DELFIA Europium Fluoroimmunoassay Fluoroimmunoassay - methods Follicle stimulating hormone Follicle Stimulating Hormone - analysis Fundamental and applied biological sciences. Psychology Fundamental immunology Humans Isotopes Molecular immunology Sensitivity and Specificity Streptavidin Techniques Thyroid stimulating hormone Thyrotropin - analysis Time Factors Time-resolved fluorometry |
| title | Europium-labelled streptavidin as a highly sensitive universal label: Indirect time-resolved immunofluorometry of FSH and TSH |
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