N008: Detection of parathyroid hypertensive factor in parathyroid cell culture media
Parathyroid hypertensive factor (PHF) is a circulating hypertensive factor found in a proportion of human essential hypertensive patients and spontaneously hypertensive rats. This study describes a new immunoassay system for detection of PHF in rat parathyroid cell culture media. We have developed a...
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| Veröffentlicht in: | American journal of hypertension 2000-04, Vol.13 (S2), p.309A-309A |
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| container_end_page | 309A |
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| container_issue | S2 |
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| container_title | American journal of hypertension |
| container_volume | 13 |
| creator | Krylova, S. Sutherland, S. Lin, M. Shan, J. Benishin, C.G. Sutherland, S.K. Lewanczuk, R.Z. |
| description | Parathyroid hypertensive factor (PHF) is a circulating hypertensive factor found in a proportion of human essential hypertensive patients and spontaneously hypertensive rats. This study describes a new immunoassay system for detection of PHF in rat parathyroid cell culture media. We have developed a hybridoma which secreted specific anti-PHF antibodies of IgM isotype. Research study demonstrated no detectable cross-reactivity with rat parathyroid hormone or shorter parathyroid hormone fragments, different vasoactive substances and plasma proteins. PHF assay is a one step competitive assay using a microtiter plates coated with monoclonal antibodies. Culture media supernatant is diluted (1:10) in a specimen diluent and incubated with horseradish peroxidase labeled PHF in microtiter plate wells. The assay has a dynamic range of 0–2.0 units/ml, with lower detection limit of 0.02 units/ml. Assay reproducibility was determined by testing three samples over the dynamic range; a precision of 14% (inter assay) and 8.3% (intra assay) was achieved. PHF immunoassay results are in agreement with previously used bioassay and show significant difference between the level of PHF secreted by parathyroid cells from spontaneously hypertensive and Wistar-Kyoto rats. In conclusion PHF enzyme immunoassay is rapid, highly sensitive and can provide a useful tool for further investigation of the physiological role of PHF. |
| doi_str_mv | 10.1016/S0895-7061(00)01104-3 |
| format | Article |
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This study describes a new immunoassay system for detection of PHF in rat parathyroid cell culture media. We have developed a hybridoma which secreted specific anti-PHF antibodies of IgM isotype. Research study demonstrated no detectable cross-reactivity with rat parathyroid hormone or shorter parathyroid hormone fragments, different vasoactive substances and plasma proteins. PHF assay is a one step competitive assay using a microtiter plates coated with monoclonal antibodies. Culture media supernatant is diluted (1:10) in a specimen diluent and incubated with horseradish peroxidase labeled PHF in microtiter plate wells. The assay has a dynamic range of 0–2.0 units/ml, with lower detection limit of 0.02 units/ml. Assay reproducibility was determined by testing three samples over the dynamic range; a precision of 14% (inter assay) and 8.3% (intra assay) was achieved. PHF immunoassay results are in agreement with previously used bioassay and show significant difference between the level of PHF secreted by parathyroid cells from spontaneously hypertensive and Wistar-Kyoto rats. In conclusion PHF enzyme immunoassay is rapid, highly sensitive and can provide a useful tool for further investigation of the physiological role of PHF.</description><identifier>ISSN: 0895-7061</identifier><identifier>EISSN: 1941-7225</identifier><identifier>DOI: 10.1016/S0895-7061(00)01104-3</identifier><identifier>CODEN: AJHYE6</identifier><language>eng</language><publisher>Oxford: Oxford University Press</publisher><subject>enzyme immunoassay ; Hypertension ; monoclonal antibody ; PHF</subject><ispartof>American journal of hypertension, 2000-04, Vol.13 (S2), p.309A-309A</ispartof><rights>Copyright Nature Publishing Group Apr 2000</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,777,781,27905,27906</link.rule.ids></links><search><creatorcontrib>Krylova, S.</creatorcontrib><creatorcontrib>Sutherland, S.</creatorcontrib><creatorcontrib>Lin, M.</creatorcontrib><creatorcontrib>Shan, J.</creatorcontrib><creatorcontrib>Benishin, C.G.</creatorcontrib><creatorcontrib>Sutherland, S.K.</creatorcontrib><creatorcontrib>Lewanczuk, R.Z.</creatorcontrib><title>N008: Detection of parathyroid hypertensive factor in parathyroid cell culture media</title><title>American journal of hypertension</title><addtitle>AJH</addtitle><description>Parathyroid hypertensive factor (PHF) is a circulating hypertensive factor found in a proportion of human essential hypertensive patients and spontaneously hypertensive rats. This study describes a new immunoassay system for detection of PHF in rat parathyroid cell culture media. We have developed a hybridoma which secreted specific anti-PHF antibodies of IgM isotype. Research study demonstrated no detectable cross-reactivity with rat parathyroid hormone or shorter parathyroid hormone fragments, different vasoactive substances and plasma proteins. PHF assay is a one step competitive assay using a microtiter plates coated with monoclonal antibodies. Culture media supernatant is diluted (1:10) in a specimen diluent and incubated with horseradish peroxidase labeled PHF in microtiter plate wells. The assay has a dynamic range of 0–2.0 units/ml, with lower detection limit of 0.02 units/ml. Assay reproducibility was determined by testing three samples over the dynamic range; a precision of 14% (inter assay) and 8.3% (intra assay) was achieved. PHF immunoassay results are in agreement with previously used bioassay and show significant difference between the level of PHF secreted by parathyroid cells from spontaneously hypertensive and Wistar-Kyoto rats. In conclusion PHF enzyme immunoassay is rapid, highly sensitive and can provide a useful tool for further investigation of the physiological role of PHF.</description><subject>enzyme immunoassay</subject><subject>Hypertension</subject><subject>monoclonal antibody</subject><subject>PHF</subject><issn>0895-7061</issn><issn>1941-7225</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><recordid>eNpVzc9LwzAYxvEgCs4ff4IQ8KKH6vs2TdJ606mbOPTgDtJLyJKUZXZtTVNx_72DieDpuXx4voScIVwhoLh-g7zgiQSBFwCXgAhZwvbICIsME5mmfJ-M_sghOer7FQBkQuCIzF8A8ht676Iz0bcNbSva6aDjchNab-ly07kQXdP7L0crbWIbqG_-EePqmpqhjkNwdO2s1yfkoNJ1705_95jMHx_m42kye508jW9niRcCkko7njNhJHeVsYs8xQIqznkqjWALnQpwRSEtmtRqbSXjOndYYJZbveDSGnZMzne3XWg_B9dHtWqH0GyLCoHlmCHLYKuSnfJ9dN-qC36tw0bp8KGEZJKr6XupSlY-i7tyosbsB8JzYp8</recordid><startdate>200004</startdate><enddate>200004</enddate><creator>Krylova, S.</creator><creator>Sutherland, S.</creator><creator>Lin, M.</creator><creator>Shan, J.</creator><creator>Benishin, C.G.</creator><creator>Sutherland, S.K.</creator><creator>Lewanczuk, R.Z.</creator><general>Oxford University Press</general><scope>BSCLL</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope></search><sort><creationdate>200004</creationdate><title>N008: Detection of parathyroid hypertensive factor in parathyroid cell culture media</title><author>Krylova, S. ; Sutherland, S. ; Lin, M. ; Shan, J. ; Benishin, C.G. ; Sutherland, S.K. ; Lewanczuk, R.Z.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-i660-fae5836c75efcdb82190f55527c63ba260e997d1c2daad735a8e19148dab57dc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>enzyme immunoassay</topic><topic>Hypertension</topic><topic>monoclonal antibody</topic><topic>PHF</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Krylova, S.</creatorcontrib><creatorcontrib>Sutherland, S.</creatorcontrib><creatorcontrib>Lin, M.</creatorcontrib><creatorcontrib>Shan, J.</creatorcontrib><creatorcontrib>Benishin, C.G.</creatorcontrib><creatorcontrib>Sutherland, S.K.</creatorcontrib><creatorcontrib>Lewanczuk, R.Z.</creatorcontrib><collection>Istex</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><jtitle>American journal of hypertension</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Krylova, S.</au><au>Sutherland, S.</au><au>Lin, M.</au><au>Shan, J.</au><au>Benishin, C.G.</au><au>Sutherland, S.K.</au><au>Lewanczuk, R.Z.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>N008: Detection of parathyroid hypertensive factor in parathyroid cell culture media</atitle><jtitle>American journal of hypertension</jtitle><addtitle>AJH</addtitle><date>2000-04</date><risdate>2000</risdate><volume>13</volume><issue>S2</issue><spage>309A</spage><epage>309A</epage><pages>309A-309A</pages><issn>0895-7061</issn><eissn>1941-7225</eissn><coden>AJHYE6</coden><abstract>Parathyroid hypertensive factor (PHF) is a circulating hypertensive factor found in a proportion of human essential hypertensive patients and spontaneously hypertensive rats. This study describes a new immunoassay system for detection of PHF in rat parathyroid cell culture media. We have developed a hybridoma which secreted specific anti-PHF antibodies of IgM isotype. Research study demonstrated no detectable cross-reactivity with rat parathyroid hormone or shorter parathyroid hormone fragments, different vasoactive substances and plasma proteins. PHF assay is a one step competitive assay using a microtiter plates coated with monoclonal antibodies. Culture media supernatant is diluted (1:10) in a specimen diluent and incubated with horseradish peroxidase labeled PHF in microtiter plate wells. The assay has a dynamic range of 0–2.0 units/ml, with lower detection limit of 0.02 units/ml. Assay reproducibility was determined by testing three samples over the dynamic range; a precision of 14% (inter assay) and 8.3% (intra assay) was achieved. PHF immunoassay results are in agreement with previously used bioassay and show significant difference between the level of PHF secreted by parathyroid cells from spontaneously hypertensive and Wistar-Kyoto rats. In conclusion PHF enzyme immunoassay is rapid, highly sensitive and can provide a useful tool for further investigation of the physiological role of PHF.</abstract><cop>Oxford</cop><pub>Oxford University Press</pub><doi>10.1016/S0895-7061(00)01104-3</doi></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0895-7061 |
| ispartof | American journal of hypertension, 2000-04, Vol.13 (S2), p.309A-309A |
| issn | 0895-7061 1941-7225 |
| language | eng |
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| source | Oxford University Press Journals All Titles (1996-Current); Alma/SFX Local Collection |
| subjects | enzyme immunoassay Hypertension monoclonal antibody PHF |
| title | N008: Detection of parathyroid hypertensive factor in parathyroid cell culture media |
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