Optimization and qualification of a functional anti-drug antibody assay for HIV-1 bnAbs

J Immunol Methods. 2020 Apr;479:112736. doi: 10.1016/j.jim.2020.112736. Epub 2020 Jan 7.


The recent identification of human monoclonal antibodies with broad and potent neutralizing activity against HIV-1 (bnAbs) has resulted in substantial efforts to develop these molecules for clinical use in the prevention and treatment of HIV-1 infection. As with any protein therapeutic drug product, it is imperative to have qualified assays that can accurately detect and quantify anti-drug antibodies (ADA) that may develop in patients receiving passive administration of HIV-1 bnAbs. Here, we have optimized and qualified a functional assay to assess the potential of ADA to inhibit the neutralizing function of HIV-1 bnAbs. Using a modified version of the validated TZM-bl HIV-1 neutralization assay, murine anti-idiotype antibodies were utilized to optimize and evaluate parameters of linearity, range, limit of detection, specificity, and precision for measuring inhibitory ADA activity against multiple HIV-1 bnAbs that are in clinical development. We further demonstrate the utility of this assay for detecting naturally occurring ADA responses in non-human primates receiving passive administration of human bnAbs. This functional assay format complements binding-antibody ADA strategies being developed for HIV-1 bnAbs, and when utilized together, will support a multi-tiered approach for ADA testing that is compliant with Good Clinical Laboratory Practice (GCLP) procedures and FDA guidance.

Keywords: Anti-drug antibody; Broadly neutralizing antibodies; HIV-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Anti-Idiotypic / analysis*
  • Antibodies, Monoclonal, Murine-Derived / analysis*
  • Broadly Neutralizing Antibodies / immunology
  • Broadly Neutralizing Antibodies / therapeutic use*
  • HIV Antibodies / immunology
  • HIV Antibodies / therapeutic use*
  • HIV Infections / therapy*
  • HIV-1 / physiology*
  • Humans
  • Mice
  • Neutralization Tests / methods*


  • Antibodies, Anti-Idiotypic
  • Antibodies, Monoclonal, Murine-Derived
  • Broadly Neutralizing Antibodies
  • HIV Antibodies