Skip to main content
Journal cover image

Optimization and validation of a neutralizing antibody assay for HIV-1 in A3R5 cells.

Publication ,  Journal Article
Sarzotti-Kelsoe, M; Daniell, X; Todd, CA; Bilska, M; Martelli, A; LaBranche, C; Perez, LG; Ochsenbauer, C; Kappes, JC; Rountree, W; Denny, TN ...
Published in: J Immunol Methods
July 2014

A3R5 is a human CD4(+) lymphoblastoid cell line that was engineered to express CCR5 and is useful for the detection of weak neutralizing antibody responses against tier 2 strains of HIV-1. Here we describe the optimization and validation of the HIV-1 neutralizing antibody assay that utilizes A3R5 cells, performed in compliance with Good Clinical Laboratory Practice (GCLP) guidelines. The assay utilizes Renilla luciferase-expressing replication competent infectious molecular clones (IMC) encoding heterologous env genes from different HIV-1 clades. Key assay validation parameters tested included specificity, accuracy, precision, limit of detection and quantitation, specificity, linearity and range, and robustness. Plasma samples demonstrated higher non-specific activity than serum samples in the A3R5 assay. This assay can tolerate a wide range of virus input but is more sensitive to cell concentration. The higher sensitivity of the A3R5 assay in neutralization responses to tier 2 strains of HIV-1 makes it complementary to, but not a substitute for the TZM-bl assay. The validated A3R5 assay is employed as an endpoint immunogenicity test for vaccine-elicited neutralizing antibodies against tier 2 strains of HIV-1, and to identify correlates of protection in HIV-1 vaccine trials conducted globally.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

July 2014

Volume

409

Start / End Page

147 / 160

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

APA
Chicago
ICMJE
MLA
NLM
Sarzotti-Kelsoe, M., Daniell, X., Todd, C. A., Bilska, M., Martelli, A., LaBranche, C., … Montefiori, D. C. (2014). Optimization and validation of a neutralizing antibody assay for HIV-1 in A3R5 cells. J Immunol Methods, 409, 147–160. https://doi.org/10.1016/j.jim.2014.02.013
Sarzotti-Kelsoe, Marcella, Xiaoju Daniell, Christopher A. Todd, Miroslawa Bilska, Amanda Martelli, Celia LaBranche, Lautaro G. Perez, et al. “Optimization and validation of a neutralizing antibody assay for HIV-1 in A3R5 cells.J Immunol Methods 409 (July 2014): 147–60. https://doi.org/10.1016/j.jim.2014.02.013.
Sarzotti-Kelsoe M, Daniell X, Todd CA, Bilska M, Martelli A, LaBranche C, et al. Optimization and validation of a neutralizing antibody assay for HIV-1 in A3R5 cells. J Immunol Methods. 2014 Jul;409:147–60.
Sarzotti-Kelsoe, Marcella, et al. “Optimization and validation of a neutralizing antibody assay for HIV-1 in A3R5 cells.J Immunol Methods, vol. 409, July 2014, pp. 147–60. Pubmed, doi:10.1016/j.jim.2014.02.013.
Sarzotti-Kelsoe M, Daniell X, Todd CA, Bilska M, Martelli A, LaBranche C, Perez LG, Ochsenbauer C, Kappes JC, Rountree W, Denny TN, Montefiori DC. Optimization and validation of a neutralizing antibody assay for HIV-1 in A3R5 cells. J Immunol Methods. 2014 Jul;409:147–160.
Journal cover image

Published In

J Immunol Methods

DOI

EISSN

1872-7905

Publication Date

July 2014

Volume

409

Start / End Page

147 / 160

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