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Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1

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.

Journal of immunological methods, 2014-07, Vol.409, p.131-146 [Periódico revisado por pares]

Netherlands: Elsevier B.V

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Título:
Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1
Autor: 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.
Assuntos: Antibodies, Neutralizing - blood ; Assay validation ; Automation, Laboratory - standards ; Biomarkers - blood ; GCLP ; Guideline Adherence - standards ; 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 ; Humans ; Limit of Detection ; Neutralization Tests - standards ; Neutralizing antibody ; Observer Variation ; Practice Guidelines as Topic - standards ; Predictive Value of Tests ; Quality Control ; Reproducibility of Results ; Time Factors ; Transfection ; TZM-bl cells
É parte de: Journal of immunological methods, 2014-07, Vol.409, p.131-146
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-3
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Descrição: 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.
Editor: Netherlands: Elsevier B.V
Idioma: Inglês

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Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1

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.

Netherlands: Elsevier B.V

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