Format

Send to

Choose Destination
J Immunol Methods. 2014 Jul;409:131-46. doi: 10.1016/j.jim.2013.11.022. Epub 2013 Dec 1.

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

Author information

1
Department of Immunology, Duke University Medical Center, Durham, NC, United States; Department of Surgery, Duke University Medical Center, Durham, NC, United States. Electronic address: marcella.sarzottikelsoe@dm.duke.edu.
2
Vaccine Research Center, National Institutes of Health (NIH), Bethesda, MD, United States.
3
Department of Surgery, Duke University Medical Center, Durham, NC, United States.
4
Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, United States.

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.

KEYWORDS:

Assay validation; GCLP; HIV; Neutralizing antibody; TZM-bl cells

PMID:
24291345
PMCID:
PMC4040342
DOI:
10.1016/j.jim.2013.11.022
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center