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

Publication ,  Journal Article
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 in: J Immunol Methods
July 2014

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.

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Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

July 2014

Volume

409

Start / End Page

131 / 146

Location

Netherlands

Related Subject Headings

  • Transfection
  • Time Factors
  • Reproducibility of Results
  • Quality Control
  • Predictive Value of Tests
  • Practice Guidelines as Topic
  • Observer Variation
  • Neutralization Tests
  • Limit of Detection
  • Immunology
 

Citation

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Sarzotti-Kelsoe, M., Bailer, R. T., Turk, E., Lin, C.-L., Bilska, M., Greene, K. M., … Montefiori, D. C. (2014). Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1. J Immunol Methods, 409, 131–146. https://doi.org/10.1016/j.jim.2013.11.022
Sarzotti-Kelsoe, Marcella, Robert T. Bailer, Ellen Turk, Chen-li Lin, Miroslawa Bilska, Kelli M. Greene, Hongmei Gao, et al. “Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1.J Immunol Methods 409 (July 2014): 131–46. https://doi.org/10.1016/j.jim.2013.11.022.
Sarzotti-Kelsoe M, Bailer RT, Turk E, Lin C-L, Bilska M, Greene KM, et al. Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1. J Immunol Methods. 2014 Jul;409:131–46.
Sarzotti-Kelsoe, Marcella, et al. “Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1.J Immunol Methods, vol. 409, July 2014, pp. 131–46. Pubmed, doi:10.1016/j.jim.2013.11.022.
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. Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1. J Immunol Methods. 2014 Jul;409:131–146.
Journal cover image

Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

July 2014

Volume

409

Start / End Page

131 / 146

Location

Netherlands

Related Subject Headings

  • Transfection
  • Time Factors
  • Reproducibility of Results
  • Quality Control
  • Predictive Value of Tests
  • Practice Guidelines as Topic
  • Observer Variation
  • Neutralization Tests
  • Limit of Detection
  • Immunology