Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1
The TZM-bl assay measures antibody-mediated neutralization of HIV-1 as a function of reductions in HIV-1 Tat-regulated firefly luciferase (Luc) reporter gene expression after a single round of infection with Env-pseudotyped viruses. This assay has become the main endpoint neutralization assay used f...
Gespeichert in:
| Veröffentlicht in: | Journal of immunological methods 2014-07, Vol.409, p.131-146 |
|---|---|
| Hauptverfasser: | , , , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 146 |
|---|---|
| container_issue | |
| container_start_page | 131 |
| container_title | Journal of immunological methods |
| container_volume | 409 |
| creator | Sarzotti-Kelsoe, Marcella Bailer, Robert T. Turk, Ellen Lin, Chen-li Bilska, Miroslawa Greene, Kelli M. Gao, Hongmei Todd, Christopher A. Ozaki, Daniel A. Seaman, Michael S. Mascola, John R. Montefiori, David C. |
| description | The TZM-bl assay measures antibody-mediated neutralization of HIV-1 as a function of reductions in HIV-1 Tat-regulated firefly luciferase (Luc) reporter gene expression after a single round of infection with Env-pseudotyped viruses. This assay has become the main endpoint neutralization assay used for the assessment of pre-clinical and clinical trial samples by a growing number of laboratories worldwide. Here we present the results of the formal optimization and validation of the TZM-bl assay, performed in compliance with Good Clinical Laboratory Practice (GCLP) guidelines. The assay was evaluated for specificity, accuracy, precision, limits of detection and quantitation, linearity, range and robustness. The validated manual TZM-bl assay was also adapted, optimized and qualified to an automated 384-well format. |
| doi_str_mv | 10.1016/j.jim.2013.11.022 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4040342</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0022175913003591</els_id><sourcerecordid>2000427859</sourcerecordid><originalsourceid>FETCH-LOGICAL-c620t-f593456c9c17fc8d95fee6c965d2d0bff553c32fd9bf7345f390766c522369813</originalsourceid><addsrcrecordid>eNqFkUFv1DAQhS0EokvhB3BBOXJJ8DixEwsJCVVAKxX1UjhwsRx7vPVqEy-2s1L31-PVlgoucLJm_M3Tm3mEvAbaAAXxbtNs_NQwCm0D0FDGnpAVDD2re0n5U7KipVVDz-UZeZHShlIKVNDn5Ix1TELb8RVZbnbZT_6gsw9zpWdb7fXW21MZXJXvsLr98bUet5VOSd9XLsQq5QLqaP0B7bGNKU0453QcmHHJsUgc_LwuetmPwXpMlV5rP6dcXV59r-Eleeb0NuGrh_ecfPv86fbisr6--XJ18fG6NoLRXDsui0lhpIHemcFK7hBLKbhllo7Ocd6aljkrR9cX0rWS9kIYzlgr5ADtOflw0t0t44TWFJPFm9pFP-l4r4L26u-f2d-pddirjna07VgRePsgEMPPBVNWk08Gt1s9Y1iSYuWmHeuHYvR_KHAhOgqDbAsKJ9TEkFJE9-gIqDomqzaqJKuOySoAVWIsM2_-XOVx4neUBXh_ArAcdO8xqmQ8zgatj2iyssH_Q_4XSvW1wg</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1566401893</pqid></control><display><type>article</type><title>Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><creator>Sarzotti-Kelsoe, Marcella ; Bailer, Robert T. ; Turk, Ellen ; Lin, Chen-li ; Bilska, Miroslawa ; Greene, Kelli M. ; Gao, Hongmei ; Todd, Christopher A. ; Ozaki, Daniel A. ; Seaman, Michael S. ; Mascola, John R. ; Montefiori, David C.</creator><creatorcontrib>Sarzotti-Kelsoe, Marcella ; Bailer, Robert T. ; Turk, Ellen ; Lin, Chen-li ; Bilska, Miroslawa ; Greene, Kelli M. ; Gao, Hongmei ; Todd, Christopher A. ; Ozaki, Daniel A. ; Seaman, Michael S. ; Mascola, John R. ; Montefiori, David C.</creatorcontrib><description>The TZM-bl assay measures antibody-mediated neutralization of HIV-1 as a function of reductions in HIV-1 Tat-regulated firefly luciferase (Luc) reporter gene expression after a single round of infection with Env-pseudotyped viruses. This assay has become the main endpoint neutralization assay used for the assessment of pre-clinical and clinical trial samples by a growing number of laboratories worldwide. Here we present the results of the formal optimization and validation of the TZM-bl assay, performed in compliance with Good Clinical Laboratory Practice (GCLP) guidelines. The assay was evaluated for specificity, accuracy, precision, limits of detection and quantitation, linearity, range and robustness. The validated manual TZM-bl assay was also adapted, optimized and qualified to an automated 384-well format.</description><identifier>ISSN: 0022-1759</identifier><identifier>EISSN: 1872-7905</identifier><identifier>DOI: 10.1016/j.jim.2013.11.022</identifier><identifier>PMID: 24291345</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Antibodies, Neutralizing - blood ; Assay validation ; Automation, Laboratory - standards ; Biomarkers - blood ; clinical trials ; compliance ; detection limit ; GCLP ; gene expression ; Guideline Adherence - standards ; guidelines ; HeLa Cells ; High-Throughput Screening Assays - standards ; HIV ; HIV Antibodies - blood ; HIV Infections - blood ; HIV Infections - diagnosis ; HIV Infections - immunology ; HIV-1 - genetics ; HIV-1 - immunology ; Human immunodeficiency virus 1 ; Humans ; Limit of Detection ; luciferase ; neutralization ; neutralization tests ; Neutralization Tests - standards ; neutralizing antibodies ; Neutralizing antibody ; Observer Variation ; Practice Guidelines as Topic - standards ; Predictive Value of Tests ; Quality Control ; reporter genes ; Reproducibility of Results ; Time Factors ; Transfection ; TZM-bl cells ; viruses</subject><ispartof>Journal of immunological methods, 2014-07, Vol.409, p.131-146</ispartof><rights>2013 Elsevier B.V.</rights><rights>Copyright © 2013 Elsevier B.V. All rights reserved.</rights><rights>2013 Elsevier B.V. All rights reserved. 2013</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c620t-f593456c9c17fc8d95fee6c965d2d0bff553c32fd9bf7345f390766c522369813</citedby><cites>FETCH-LOGICAL-c620t-f593456c9c17fc8d95fee6c965d2d0bff553c32fd9bf7345f390766c522369813</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0022175913003591$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,776,780,881,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24291345$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sarzotti-Kelsoe, Marcella</creatorcontrib><creatorcontrib>Bailer, Robert T.</creatorcontrib><creatorcontrib>Turk, Ellen</creatorcontrib><creatorcontrib>Lin, Chen-li</creatorcontrib><creatorcontrib>Bilska, Miroslawa</creatorcontrib><creatorcontrib>Greene, Kelli M.</creatorcontrib><creatorcontrib>Gao, Hongmei</creatorcontrib><creatorcontrib>Todd, Christopher A.</creatorcontrib><creatorcontrib>Ozaki, Daniel A.</creatorcontrib><creatorcontrib>Seaman, Michael S.</creatorcontrib><creatorcontrib>Mascola, John R.</creatorcontrib><creatorcontrib>Montefiori, David C.</creatorcontrib><title>Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>The TZM-bl assay measures antibody-mediated neutralization of HIV-1 as a function of reductions in HIV-1 Tat-regulated firefly luciferase (Luc) reporter gene expression after a single round of infection with Env-pseudotyped viruses. This assay has become the main endpoint neutralization assay used for the assessment of pre-clinical and clinical trial samples by a growing number of laboratories worldwide. Here we present the results of the formal optimization and validation of the TZM-bl assay, performed in compliance with Good Clinical Laboratory Practice (GCLP) guidelines. The assay was evaluated for specificity, accuracy, precision, limits of detection and quantitation, linearity, range and robustness. The validated manual TZM-bl assay was also adapted, optimized and qualified to an automated 384-well format.</description><subject>Antibodies, Neutralizing - blood</subject><subject>Assay validation</subject><subject>Automation, Laboratory - standards</subject><subject>Biomarkers - blood</subject><subject>clinical trials</subject><subject>compliance</subject><subject>detection limit</subject><subject>GCLP</subject><subject>gene expression</subject><subject>Guideline Adherence - standards</subject><subject>guidelines</subject><subject>HeLa Cells</subject><subject>High-Throughput Screening Assays - standards</subject><subject>HIV</subject><subject>HIV Antibodies - blood</subject><subject>HIV Infections - blood</subject><subject>HIV Infections - diagnosis</subject><subject>HIV Infections - immunology</subject><subject>HIV-1 - genetics</subject><subject>HIV-1 - immunology</subject><subject>Human immunodeficiency virus 1</subject><subject>Humans</subject><subject>Limit of Detection</subject><subject>luciferase</subject><subject>neutralization</subject><subject>neutralization tests</subject><subject>Neutralization Tests - standards</subject><subject>neutralizing antibodies</subject><subject>Neutralizing antibody</subject><subject>Observer Variation</subject><subject>Practice Guidelines as Topic - standards</subject><subject>Predictive Value of Tests</subject><subject>Quality Control</subject><subject>reporter genes</subject><subject>Reproducibility of Results</subject><subject>Time Factors</subject><subject>Transfection</subject><subject>TZM-bl cells</subject><subject>viruses</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUFv1DAQhS0EokvhB3BBOXJJ8DixEwsJCVVAKxX1UjhwsRx7vPVqEy-2s1L31-PVlgoucLJm_M3Tm3mEvAbaAAXxbtNs_NQwCm0D0FDGnpAVDD2re0n5U7KipVVDz-UZeZHShlIKVNDn5Ix1TELb8RVZbnbZT_6gsw9zpWdb7fXW21MZXJXvsLr98bUet5VOSd9XLsQq5QLqaP0B7bGNKU0453QcmHHJsUgc_LwuetmPwXpMlV5rP6dcXV59r-Eleeb0NuGrh_ecfPv86fbisr6--XJ18fG6NoLRXDsui0lhpIHemcFK7hBLKbhllo7Ocd6aljkrR9cX0rWS9kIYzlgr5ADtOflw0t0t44TWFJPFm9pFP-l4r4L26u-f2d-pddirjna07VgRePsgEMPPBVNWk08Gt1s9Y1iSYuWmHeuHYvR_KHAhOgqDbAsKJ9TEkFJE9-gIqDomqzaqJKuOySoAVWIsM2_-XOVx4neUBXh_ArAcdO8xqmQ8zgatj2iyssH_Q_4XSvW1wg</recordid><startdate>20140701</startdate><enddate>20140701</enddate><creator>Sarzotti-Kelsoe, Marcella</creator><creator>Bailer, Robert T.</creator><creator>Turk, Ellen</creator><creator>Lin, Chen-li</creator><creator>Bilska, Miroslawa</creator><creator>Greene, Kelli M.</creator><creator>Gao, Hongmei</creator><creator>Todd, Christopher A.</creator><creator>Ozaki, Daniel A.</creator><creator>Seaman, Michael S.</creator><creator>Mascola, John R.</creator><creator>Montefiori, David C.</creator><general>Elsevier B.V</general><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><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20140701</creationdate><title>Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1</title><author>Sarzotti-Kelsoe, Marcella ; Bailer, Robert T. ; Turk, Ellen ; Lin, Chen-li ; Bilska, Miroslawa ; Greene, Kelli M. ; Gao, Hongmei ; Todd, Christopher A. ; Ozaki, Daniel A. ; Seaman, Michael S. ; Mascola, John R. ; Montefiori, David C.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c620t-f593456c9c17fc8d95fee6c965d2d0bff553c32fd9bf7345f390766c522369813</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Antibodies, Neutralizing - blood</topic><topic>Assay validation</topic><topic>Automation, Laboratory - standards</topic><topic>Biomarkers - blood</topic><topic>clinical trials</topic><topic>compliance</topic><topic>detection limit</topic><topic>GCLP</topic><topic>gene expression</topic><topic>Guideline Adherence - standards</topic><topic>guidelines</topic><topic>HeLa Cells</topic><topic>High-Throughput Screening Assays - standards</topic><topic>HIV</topic><topic>HIV Antibodies - blood</topic><topic>HIV Infections - blood</topic><topic>HIV Infections - diagnosis</topic><topic>HIV Infections - immunology</topic><topic>HIV-1 - genetics</topic><topic>HIV-1 - immunology</topic><topic>Human immunodeficiency virus 1</topic><topic>Humans</topic><topic>Limit of Detection</topic><topic>luciferase</topic><topic>neutralization</topic><topic>neutralization tests</topic><topic>Neutralization Tests - standards</topic><topic>neutralizing antibodies</topic><topic>Neutralizing antibody</topic><topic>Observer Variation</topic><topic>Practice Guidelines as Topic - standards</topic><topic>Predictive Value of Tests</topic><topic>Quality Control</topic><topic>reporter genes</topic><topic>Reproducibility of Results</topic><topic>Time Factors</topic><topic>Transfection</topic><topic>TZM-bl cells</topic><topic>viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sarzotti-Kelsoe, Marcella</creatorcontrib><creatorcontrib>Bailer, Robert T.</creatorcontrib><creatorcontrib>Turk, Ellen</creatorcontrib><creatorcontrib>Lin, Chen-li</creatorcontrib><creatorcontrib>Bilska, Miroslawa</creatorcontrib><creatorcontrib>Greene, Kelli M.</creatorcontrib><creatorcontrib>Gao, Hongmei</creatorcontrib><creatorcontrib>Todd, Christopher A.</creatorcontrib><creatorcontrib>Ozaki, Daniel A.</creatorcontrib><creatorcontrib>Seaman, Michael S.</creatorcontrib><creatorcontrib>Mascola, John R.</creatorcontrib><creatorcontrib>Montefiori, David C.</creatorcontrib><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><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sarzotti-Kelsoe, Marcella</au><au>Bailer, Robert T.</au><au>Turk, Ellen</au><au>Lin, Chen-li</au><au>Bilska, Miroslawa</au><au>Greene, Kelli M.</au><au>Gao, Hongmei</au><au>Todd, Christopher A.</au><au>Ozaki, Daniel A.</au><au>Seaman, Michael S.</au><au>Mascola, John R.</au><au>Montefiori, David C.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>2014-07-01</date><risdate>2014</risdate><volume>409</volume><spage>131</spage><epage>146</epage><pages>131-146</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><abstract>The TZM-bl assay measures antibody-mediated neutralization of HIV-1 as a function of reductions in HIV-1 Tat-regulated firefly luciferase (Luc) reporter gene expression after a single round of infection with Env-pseudotyped viruses. This assay has become the main endpoint neutralization assay used for the assessment of pre-clinical and clinical trial samples by a growing number of laboratories worldwide. Here we present the results of the formal optimization and validation of the TZM-bl assay, performed in compliance with Good Clinical Laboratory Practice (GCLP) guidelines. The assay was evaluated for specificity, accuracy, precision, limits of detection and quantitation, linearity, range and robustness. The validated manual TZM-bl assay was also adapted, optimized and qualified to an automated 384-well format.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>24291345</pmid><doi>10.1016/j.jim.2013.11.022</doi><tpages>16</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0022-1759 |
| ispartof | Journal of immunological methods, 2014-07, Vol.409, p.131-146 |
| issn | 0022-1759 1872-7905 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4040342 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Antibodies, Neutralizing - blood Assay validation Automation, Laboratory - standards Biomarkers - blood clinical trials compliance detection limit GCLP gene expression Guideline Adherence - standards guidelines HeLa Cells High-Throughput Screening Assays - standards HIV HIV Antibodies - blood HIV Infections - blood HIV Infections - diagnosis HIV Infections - immunology HIV-1 - genetics HIV-1 - immunology Human immunodeficiency virus 1 Humans Limit of Detection luciferase neutralization neutralization tests Neutralization Tests - standards neutralizing antibodies Neutralizing antibody Observer Variation Practice Guidelines as Topic - standards Predictive Value of Tests Quality Control reporter genes Reproducibility of Results Time Factors Transfection TZM-bl cells viruses |
| title | Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1 |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-07T20%3A01%3A57IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Optimization%20and%20validation%20of%20the%20TZM-bl%20assay%20for%20standardized%20assessments%20of%20neutralizing%20antibodies%20against%20HIV-1&rft.jtitle=Journal%20of%20immunological%20methods&rft.au=Sarzotti-Kelsoe,%20Marcella&rft.date=2014-07-01&rft.volume=409&rft.spage=131&rft.epage=146&rft.pages=131-146&rft.issn=0022-1759&rft.eissn=1872-7905&rft_id=info:doi/10.1016/j.jim.2013.11.022&rft_dat=%3Cproquest_pubme%3E2000427859%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1566401893&rft_id=info:pmid/24291345&rft_els_id=S0022175913003591&rfr_iscdi=true |