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

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Sarzotti-Kelsoe, M; Bailer, RT; Turk, E; Lin, C-L; Bilska, M; Greene, KM; Gao, H; Todd, CA; Ozaki, DA; Seaman, MS; Mascola, JR; Montefiori, DC

Published Date

  • July 2014

Published In

Volume / Issue

  • 409 /

Start / End Page

  • 131 - 146

PubMed ID

  • 24291345

Pubmed Central ID

  • 24291345

Electronic International Standard Serial Number (EISSN)

  • 1872-7905

Digital Object Identifier (DOI)

  • 10.1016/j.jim.2013.11.022

Language

  • eng

Conference Location

  • Netherlands