A humanized nanobody phage display library yields potent binders of SARS CoV-2 spike

Abstract

Neutralizing antibodies targeting the SARS-CoV-2 spike protein have shown a great preventative/therapeutic potential. Here, we report a rapid and efficient strategy for the development and design of SARS-CoV-2 neutralizing humanized nanobody constructs with sub-nanomolar affinities and nanomolar potencies. CryoEM-based structural analysis of the nanobodies in complex with spike revealed two distinct binding modes. The most potent nanobody, RBD-1-2G(NCATS-BL8125), tolerates the N501Y RBD mutation and remains capable of neutralizing the B.1.1.7 (Alpha) variant. Molecular dynamics simulations provide a structural basis for understanding the neutralization process of nanobodies exclusively focused on the spike-ACE2 interface with and without the N501Y mutation on RBD. A primary human airway air-lung interface (ALI) ex vivo model showed that RBD-1-2G-Fc antibody treatment was effective at reducing viral burden following WA1 and B.1.1.7 SARS-CoV-2 infections. Therefore, this presented strategy will serve as a tool to mitigate the threat of emerging SARS-CoV-2 variants.

Document Details

Document Type
Pub Defense Publication
Publication Date
Aug 10, 2022
Source ID
10.1371/journal.pone.0272364

Entities

People

  • Alex Renn
  • Allen L Hsu
  • Anton Simeonov
  • Bryan D Fleming
  • Catherine Z. Chen
  • Dominic Esposito
  • Emily M. Lee
  • Eunkeu Oh
  • Juliana Da Fonseca Rezende E Mello
  • Kedar Sharma
  • Kimihiro Susumu
  • Kirill Gorshkov
  • Lalith Perera
  • Manisha Pradhan
  • Marc Ferrer
  • Mario J. Borgnia
  • Mason Wolak
  • Matthew D. Hall
  • Miao Xu
  • Nitya Ramakrishnan
  • Quinlin Hanson
  • Richard T. Eastman
  • Robert Petrovich
  • Shelley Perkins
  • Thomas B. Stanley
  • Troy Taylor
  • Venkata Dandey
  • Wei Zheng
  • William Gillette
  • Xin Hu
  • Ying Fu
  • Zina Itkin

Organizations

  • Istituto Superiore di Sanità
  • National Cancer Institute
  • National Center for Advancing Translational Sciences
  • National Institute of Environmental Health Sciences
  • United States Naval Research Laboratory

Tags

Fields of Study

  • Biology

Readers

  • Immunology
  • Molecular Genetics
  • Pulsed Power and Plasma Physics.