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Related Experiment Video

Updated: Nov 17, 2025

Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays
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Published on: September 26, 2011

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A sensitive method to quantify HIV-1 antibodies in mucosal samples.

Madhu Prabhakaran1, Sandeep Narpala1, Sarah F Andrews1

  • 1Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 40 Convent Drive, Bethesda, MD 20814, USA.

Journal of Immunological Methods
|February 14, 2021
PubMed
Summary

A new ultra-sensitive assay accurately measures broadly neutralizing antibodies (bNabs) in mucosal samples, crucial for HIV prevention research. This advance aids in understanding antibody effectiveness and optimizing future HIV vaccine trials.

Keywords:
Human immunodeficiency virus (HIV)Monoclonal antibodiesMucosal tissuesSingulexUltra-sensitive quantificationVRC01

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Area of Science:

  • Immunology
  • Virology
  • Analytical Chemistry

Background:

  • Human immunodeficiency virus (HIV) remains a global health challenge.
  • Passive immunization using broadly neutralizing antibodies (bNabs) is a promising HIV intervention strategy.
  • VRC01 and its variants are key bNab candidates evaluated in clinical trials for HIV prevention and treatment.

Purpose of the Study:

  • To develop and validate an ultra-sensitive immunoassay for quantifying VRC01, VRC01-LS, VRC07, and VRC07-523LS.
  • To measure bNab concentrations in mucosal compartments (rectal, cervical, oral) for pharmacokinetic and bioavailability analyses.
  • To assess the assay's performance against traditional methods and its utility in clinical trial sample analysis.

Main Methods:

  • Development of a novel immunoassay on the Singulex platform for bNab quantification.
  • Assay validation demonstrating a >4-log linear dynamic range and <120 pg/mL lower limit of quantitation (LLOQ).
  • Application of the assay to quantify VRC01 in mucosal and serum samples from VRC 601 and VRC 602 clinical trials.

Main Results:

  • The developed assay achieved ultra-sensitive quantification of bNabs in mucosal samples.
  • VRC01 levels in a significant proportion of oral (37%) and rectal (25%) mucosal samples were below the LLOQ of traditional immunoassays.
  • The assay showed excellent correlation (r²=0.9509) with ELISA for serum VRC01 measurements.

Conclusions:

  • The novel immunoassay provides a sensitive and accurate method for quantifying bNabs in complex biological matrices like mucosal tissues.
  • This assay facilitates the determination of antibody infiltration and bioavailability, essential for understanding HIV protection.
  • The findings will inform dosing regimens and clinical trial designs for future bNab-based HIV prevention strategies.