Optimization and validation of a neutralizing antibody assay for HIV-1 in A3R5 cells.
| dc.contributor.author | Sarzotti-Kelsoe, M | |
| dc.contributor.author | Daniell, X | |
| dc.contributor.author | Todd, CA | |
| dc.contributor.author | Bilska, M | |
| dc.contributor.author | Martelli, A | |
| dc.contributor.author | LaBranche, C | |
| dc.contributor.author | Perez, LG | |
| dc.contributor.author | Ochsenbauer, C | |
| dc.contributor.author | Kappes, JC | |
| dc.contributor.author | Rountree, W | |
| dc.contributor.author | Denny, TN | |
| dc.contributor.author | Montefiori, DC | |
| dc.coverage.spatial | Netherlands | |
| dc.date.accessioned | 2017-06-01T19:27:37Z | |
| dc.date.available | 2017-06-01T19:27:37Z | |
| dc.date.issued | 2014-07 | |
| dc.description.abstract | 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. | |
| dc.identifier | ||
| dc.identifier | S0022-1759(14)00072-6 | |
| dc.identifier.eissn | 1872-7905 | |
| dc.identifier.uri | ||
| dc.language | eng | |
| dc.publisher | Springer Science and Business Media LLC | |
| dc.relation.ispartof | J Immunol Methods | |
| dc.relation.isversionof | 10.1016/j.jim.2014.02.013 | |
| dc.subject | A3R5 cells | |
| dc.subject | Assay validation | |
| dc.subject | HIV | |
| dc.subject | Neutralizing antibody | |
| dc.subject | Antibodies, Neutralizing | |
| dc.subject | Automation, Laboratory | |
| dc.subject | Biomarkers | |
| dc.subject | Cell Line | |
| dc.subject | Guideline Adherence | |
| dc.subject | HIV Antibodies | |
| dc.subject | HIV Infections | |
| dc.subject | HIV-1 | |
| dc.subject | High-Throughput Screening Assays | |
| dc.subject | Humans | |
| dc.subject | Limit of Detection | |
| dc.subject | Neutralization Tests | |
| dc.subject | Observer Variation | |
| dc.subject | Practice Guidelines as Topic | |
| dc.subject | Predictive Value of Tests | |
| dc.subject | Quality Control | |
| dc.subject | Reproducibility of Results | |
| dc.subject | Time Factors | |
| dc.subject | Transfection | |
| dc.title | Optimization and validation of a neutralizing antibody assay for HIV-1 in A3R5 cells. | |
| dc.type | Journal article | |
| duke.contributor.orcid | LaBranche, C|0000-0002-6653-6655 | |
| duke.contributor.orcid | Montefiori, DC|0000-0003-0856-6319 | |
| pubs.author-url | ||
| pubs.begin-page | 147 | |
| pubs.end-page | 160 | |
| pubs.organisational-group | Basic Science Departments | |
| pubs.organisational-group | Clinical Science Departments | |
| pubs.organisational-group | Duke | |
| pubs.organisational-group | Duke Human Vaccine Institute | |
| pubs.organisational-group | Immunology | |
| pubs.organisational-group | Institutes and Centers | |
| pubs.organisational-group | Medicine | |
| pubs.organisational-group | Medicine, Duke Human Vaccine Institute | |
| pubs.organisational-group | School of Medicine | |
| pubs.organisational-group | Surgery | |
| pubs.organisational-group | Surgery, Surgical Sciences | |
| pubs.publication-status | Published | |
| pubs.volume | 409 |