Immunogenicity Risk Profile of Nanobodies

Front Immunol. 2021 Mar 9;12:632687. doi: 10.3389/fimmu.2021.632687. eCollection 2021.


Nanobodies (Nbs), the variable domains of camelid heavy chain-only antibodies, are a promising class of therapeutics or in vivo imaging reagents entering the clinic. They possess unique characteristics, including a minimal size, providing fast pharmacokinetics, high-target specificity, and an affinity in the (sub-)nanomolar range in conjunction with an easy selection and production, which allow them to outperform conventional antibodies for imaging and radiotherapeutic purposes. As for all protein theranostics, extended safety assessment and investigation of their possible immunogenicity in particular are required. In this study, we assessed the immunogenicity risk profile of two Nbs that are in phase II clinical trials: a first Nb against Human Epidermal growth factor Receptor 2 (HER2) for PET imaging of breast cancer and a second Nb with specificity to the Macrophage Mannose Receptor (MMR) for PET imaging of tumor-associated macrophages. For the anti-HER2 Nb, we show that only one out of 20 patients had a low amount of pre-existing anti-drug antibodies (ADAs), which only marginally increased 3 months after administering the Nb, and without negative effects of safety and pharmacokinetics. Further in vitro immunogenicity assessment assays showed that both non-humanized Nbs were taken up by human dendritic cells but exhibited no or only a marginal capacity to activate dendritic cells or to induce T cell proliferation. From our data, we conclude that monomeric Nbs present a low immunogenicity risk profile, which is encouraging for their future development toward potential clinical applications.

One sentence summary: Nanobodies, the recombinant single domain affinity reagents derived from heavy chain-only antibodies in camelids, are proven to possess a low immunogenicity risk profile, which will facilitate a growing number of Nanobodies to enter the clinic for therapeutic or in vivo diagnostic applications.

Keywords: DC activation; T cell—DC interactions; anti drug antibodies; dendritic cells; immunogenicity; nanobody.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / blood
  • Camelids, New World
  • Cell Proliferation
  • Coculture Techniques
  • Cytokines / metabolism
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism
  • Humans
  • Membrane Glycoproteins / immunology
  • Receptor, ErbB-2 / immunology
  • Receptors, Immunologic / immunology
  • Single-Domain Antibodies / administration & dosage
  • Single-Domain Antibodies / immunology*
  • T-Lymphocytes / immunology


  • Antibodies
  • Cytokines
  • MRC1 protein, human
  • Membrane Glycoproteins
  • Receptors, Immunologic
  • Single-Domain Antibodies
  • ERBB2 protein, human
  • Receptor, ErbB-2