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

Jennifer M Zupancic; John S Schardt; Alec A Desai; Emily K Makowski; Matthew D Smith; Ghasidit Pornnoppadol; Mayara Garcia de Mattos Barbosa; Marilia Cascalho; Thomas M Lanigan; Peter M Tessier

doi:10.1002/adtp.202100099

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

Adv Ther (Weinh). 2021 Aug;4(8):2100099. doi: 10.1002/adtp.202100099. Epub 2021 Aug 2.

Authors

Jennifer M Zupancic^{1

2}, John S Schardt^{1

2

3}, Alec A Desai^{1

2}, Emily K Makowski^{2

3}, Matthew D Smith^{1

2}, Ghasidit Pornnoppadol^{2

3}, Mayara Garcia de Mattos Barbosa⁴, Marilia Cascalho^{4

5}, Thomas M Lanigan⁶, Peter M Tessier^{1

2

3

7}

Affiliations

¹ Department of Chemical Engineering University of Michigan Ann Arbor MI 48109 USA.
² Biointerfaces Institute University of Michigan Ann Arbor MI 48109 USA.
³ Department of Pharmaceutical Sciences University of Michigan Ann Arbor MI 48109 USA.
⁴ Department of Surgery University of Michigan Ann Arbor MI 48109 USA.
⁵ Department of Microbiology and Immunology University of Michigan Ann Arbor MI 48109 USA.
⁶ Division of Rheumatology Department of Internal Medicine University of Michigan Medical School Ann Arbor MI 48109 USA.
⁷ Department of Biomedical Engineering University of Michigan Ann Arbor MI 48109 USA.

Abstract

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.

Keywords: COVID‐19; antibody; antibody fragment; camelid; polyvalency; polyvalent; protein engineering.

Abstract

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