Engineered Ultra-High Affinity Synthetic Antibodies for SARS-CoV-2 Neutralization and Detection

J Mol Biol. 2021 May 14;433(10):166956. doi: 10.1016/j.jmb.2021.166956. Epub 2021 Mar 26.

Abstract

The Covid-19 pandemic is a centenarial global catastrophe. Similar events are likely to be recurring with more frequency in the future. The inability to control the virus' impact is caused by many factors, but the lack of a technology infrastructure to detect and impede the virus at an early stage are principal shortcomings. Using phage display mutagenesis, we have generated a cohort of high performance antibody fragments (Fabs) that can be used in a sensitive point of care (POC) assay and are potent inhibitors (IC50-0.5 nM) to viral entry into cells. The POC assay is based on a split-enzyme (β-lactamase) complementation strategy that detects virus particles at low nM levels. We have shown that this assay is equally effective for detecting other viruses like Ebola and Zika. Importantly, its components can be freeze dried and stored, but becomes fully active when rehydrated.

Keywords: Covid-19; antiviral therapeutics; point-of-care virus detection; protein engineering; split-enzyme complementation assay.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Neutralizing / chemistry*
  • Antibodies, Neutralizing / immunology*
  • Antibody Affinity*
  • Chlorocebus aethiops
  • Epitopes, B-Lymphocyte / chemistry
  • Epitopes, B-Lymphocyte / immunology
  • Freeze Drying
  • Genetic Complementation Test
  • Immunoglobulin Fab Fragments / chemistry*
  • Immunoglobulin Fab Fragments / immunology*
  • Neutralization Tests
  • Peptide Library
  • Point-of-Care Systems
  • SARS-CoV-2 / immunology*
  • SARS-CoV-2 / isolation & purification*
  • Spike Glycoprotein, Coronavirus / chemistry
  • Spike Glycoprotein, Coronavirus / immunology
  • Vero Cells
  • Virus Internalization
  • beta-Lactamases / metabolism

Substances

  • Antibodies, Neutralizing
  • Epitopes, B-Lymphocyte
  • Immunoglobulin Fab Fragments
  • Peptide Library
  • Spike Glycoprotein, Coronavirus
  • spike protein, SARS-CoV-2
  • beta-Lactamases