High affinity nanobodies block SARS-CoV-2 spike receptor binding domain interaction with human angiotensin converting enzyme

Thomas J Esparza; Negin P Martin; George P Anderson; Ellen R Goldman; David L Brody

doi:10.1038/s41598-020-79036-0

High affinity nanobodies block SARS-CoV-2 spike receptor binding domain interaction with human angiotensin converting enzyme

Sci Rep. 2020 Dec 22;10(1):22370. doi: 10.1038/s41598-020-79036-0.

Authors

Thomas J Esparza^{1

2}, Negin P Martin^{3

4}, George P Anderson⁵, Ellen R Goldman⁵, David L Brody^{6

7}

Affiliations

¹ Laboratory of Functional and Molecular Imaging, The National Institute of Neurological Disorders and Stroke Intramural Research Program, Bethesda, MD, 20892, USA.
² Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, 20892, USA.
³ Viral Vector Core, National Institute of Environmental Health Sciences, NIH/DHHS, Research Triangle Park, NC, 27709, USA.
⁴ Neurobiology Laboratory, National Institute of Environmental Health Sciences, NIH/DHHS, Research Triangle Park, NC, 27709, USA.
⁵ Center for Biomolecular Science and Engineering, US Naval Research Laboratory, Washington, DC, 20375, USA.
⁶ Laboratory of Functional and Molecular Imaging, The National Institute of Neurological Disorders and Stroke Intramural Research Program, Bethesda, MD, 20892, USA. david.brody@nih.gov.
⁷ Department of Neurology, Uniformed Services University of the Health Sciences, Bethesda, MD, 20814, USA. david.brody@nih.gov.

Abstract

There are currently few approved effective treatments for SARS-CoV-2, the virus responsible for the COVID-19 pandemic. Nanobodies are 12-15 kDa single-domain antibody fragments that can be delivered by inhalation and are amenable to relatively inexpensive large scale production compared to other biologicals. We have isolated nanobodies that bind to the SARS-CoV-2 spike protein receptor binding domain and block spike protein interaction with the angiotensin converting enzyme 2 (ACE2) with 1-5 nM affinity. The lead nanobody candidate, NIH-CoVnb-112, blocks SARS-CoV-2 spike pseudotyped lentivirus infection of HEK293 cells expressing human ACE2 with an EC₅₀ of 0.3 µg/mL. NIH-CoVnb-112 retains structural integrity and potency after nebulization. Furthermore, NIH-CoVnb-112 blocks interaction between ACE2 and several high affinity variant forms of the spike protein. These nanobodies and their derivatives have therapeutic, preventative, and diagnostic potential.

Publication types

Research Support, N.I.H., Intramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Angiotensin-Converting Enzyme 2 / genetics
Angiotensin-Converting Enzyme 2 / metabolism*
Animals
Antibodies, Neutralizing / immunology
Antibodies, Viral / immunology*
Antibody Affinity*
Binding Sites, Antibody / immunology
COVID-19 / metabolism*
COVID-19 / therapy
COVID-19 / virology
Camelids, New World
Drug Discovery / methods*
HEK293 Cells
Humans
Immunization / methods
Male
Protein Binding
Protein Interaction Domains and Motifs / immunology*
SARS-CoV-2 / chemistry*
Signal Transduction / genetics
Single-Domain Antibodies / immunology*
Spike Glycoprotein, Coronavirus / chemistry*
Spike Glycoprotein, Coronavirus / genetics
Transduction, Genetic
Transfection

Substances

Antibodies, Neutralizing
Antibodies, Viral
Single-Domain Antibodies
Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2
ACE2 protein, human
Angiotensin-Converting Enzyme 2