Immunogenicity and humanization of single‐domain antibodies

Rossotti, Martín A.; Bélanger, Kasandra; Henry, Kevin A.; Tanha, Jamshid

doi:10.1111/febs.15809

Cited by 94 publications

(72 citation statements)

References 140 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Inspired by the FDA approval of the first Nb therapy (Morrison, 2019), a number of preclinical and clinical programs have been initiated, providing critical insights into the safety profiles, dosing regime, and efficacy of therapeutic Nbs. For example, recent studies based on dozens of preclinical results found that ADA responses were minor and were generally non-neutralizing to compromise efficacy (Rossotti et al, 2021). Clinical trials have further revealed limited safety issues of Nbs (mostly humanized Nbs) in both patients and healthy volunteers.…”

Section: Discussionmentioning

confidence: 99%

Llamanade: An open-source computational pipeline for robust nanobody humanization

et al. 2022

View full text Add to dashboard Cite

Nanobodies (Nbs) have emerged as a promising class of biologics. Despite having marked physicochemical properties, Nbs are derived from camelids and may require humanization to improve translational potentials. By systematically analyzing the sequence and structural properties of Nbs, we found substantial framework diversities and revealed the key differences between Nbs and human immunoglobulin G antibodies. We identified conserved residues that may contribute to enhanced solubility, structural stability, and antigen binding, providing insights into Nb humanization. Based on big data analysis, we developed "Llamanade," an opensource software to facilitate rational humanization of Nbs. Using sequence as input, Llamanade can rapidly extract sequence features, model structures, and optimize solutions to humanize Nbs. Finally, we used Llamanade to successfully humanize a cohort of structurally diverse and potent SARS-CoV-2 neutralizing Nbs. Llamanade is freely available and will be easily accessible on a server to support the development of therapeutic Nbs into safe and effective trials.

show abstract

Section: Discussionmentioning

confidence: 99%

Llamanade: An open-source computational pipeline for robust nanobody humanization

et al. 2022

View full text Add to dashboard Cite

show abstract

“… 44 The ACE2 ECD was fused to either the N- or C-terminus of the Fc while the anti-pIgR VHH moieties were formatted only on the N-terminus of the bifunctional molecules to avoid binding with preformed antibodies in human sera. 45 All molecules featured a human IgG1-based constant region.…”

Section: Resultsmentioning

confidence: 99%

Bifunctional molecules targeting SARS-CoV-2 spike and the polymeric Ig receptor display neutralization activity and mucosal enrichment

White

Tamot

Doddareddy

et al. 2021

mAbs

View full text Add to dashboard Cite

The global health crisis and economic tolls of COVID-19 necessitate a panoply of strategies to treat SARS-CoV-2 infection. To date, few treatment options exist, although neutralizing antibodies against the spike glycoprotein have proven to be effective. Because infection is initiated at the mucosa and propagates mainly at this site throughout the course of the disease, blocking the virus at the mucosal milieu should be effective. However, administration of biologics to the mucosa presents a substantial challenge. Here, we describe bifunctional molecules combining single-domain variable regions that bind to the polymeric Ig receptor (pIgR) and to the SARS-CoV-2 spike protein via addition of the ACE2 extracellular domain (ECD). The hypothesis behind this design is that pIgR will transport the molecule from the circulation to the mucosal surface where the ACE ECD would act as a decoy receptor for the nCoV2. The bifunctional molecules bind SARS-Cov-2 spike glycoprotein in vitro and efficiently transcytose across the lung epithelium in human tissue-based analyses. Designs featuring ACE2 tethered to the C-terminus of the Fc do not induce antibody-dependent cytotoxicity against pIgR-expressing cells. These molecules thus represent a potential therapeutic modality for systemic administration of neutralizing anti-SARS-CoV-2 molecules to the mucosa.

show abstract

“…However, camelid antibodies have been reported to share greater sequence similarity with human antibodies than with antibodies from other commonly immunized species, such as mice. [ 47 ] While immunogenicity of nanobodies is difficult to predict and direct analysis is beyond the scope of this study, nanobodies have been successfully humanized and administered as therapeutics. The first therapeutic nanobody, caplacizumab, was approved by the FDA in 2019, [ 48 ] demonstrating the potential of nanobodies in therapeutic applications.…”

Section: Discussionmentioning

confidence: 99%

Engineered Multivalent Nanobodies Potently and Broadly Neutralize SARS‐CoV‐2 Variants

Zupancic

Schardt

Desai

et al. 2021

Advanced Therapeutics

View full text Add to dashboard Cite

The COVID-19 pandemic continues to be a severe threat to human health, especially due to current and emerging SARS-CoV-2 variants with potential to escape humoral immunity developed after vaccination or infection. The development of broadly neutralizing antibodies that engage evolutionarily conserved epitopes on coronavirus spike proteins represents a promising strategy to improve therapy and prophylaxis against SARS-CoV-2 and variants thereof. Herein, a facile multivalent engineering approach is employed to achieve large synergistic improvements in the neutralizing activity of a SARS-CoV-2 cross-reactive nanobody (VHH-72) initially generated against SARS-CoV. This synergy is epitope specific and is not observed for a second high-affinity nanobody against a non-conserved epitope in the receptor-binding domain. Importantly, a hexavalent VHH-72 nanobody retains binding to spike proteins from multiple highly transmissible SARS-CoV-2 variants (B.1.1.7 and B.1.351) and potently neutralizes them. Multivalent VHH-72 nanobodies also display drug-like biophysical properties, including high stability, high solubility, and low levels of non-specific binding. The unique neutralizing and biophysical properties of VHH-72 multivalent nanobodies make them attractive as therapeutics against SARS-CoV-2 variants.

show abstract

Immunogenicity and humanization of single‐domain antibodies

Abstract: receptor 1; VEGF, vascular endothelial growth factor; V H , variable heavy chain domain; V L , variable light chain domain; V H H, autonomous variable domain of camelid heavy chain-only antibody; V NAR , autonomous variable domain of shark Ig new antigen receptor.

Cited by 94 publications

References 140 publications

Llamanade: An open-source computational pipeline for robust nanobody humanization

Llamanade: An open-source computational pipeline for robust nanobody humanization

Bifunctional molecules targeting SARS-CoV-2 spike and the polymeric Ig receptor display neutralization activity and mucosal enrichment

Engineered Multivalent Nanobodies Potently and Broadly Neutralize SARS‐CoV‐2 Variants

Contact Info

Product

Resources

About