Rapid discovery and optimization of therapeutic antibodies against emerging infectious diseases

Protein Eng Des Sel. 2008 Aug;21(8):495-505. doi: 10.1093/protein/gzn027. Epub 2008 May 13.

Abstract

Using a comprehensive set of discovery and optimization tools, antibodies were produced with the ability to neutralize SARS coronavirus (SARS-CoV) infection in Vero E6 cells and in animal models. These anti-SARS antibodies were discovered using a novel DNA display method, which can identify new antibodies within days. Once neutralizing antibodies were identified, a comprehensive and effective means of converting the mouse sequences to human frameworks was accomplished using HuFR (human framework reassembly) technology. The best variant (61G4) from this screen showed a 3.5-4-fold improvement in neutralization of SARS-CoV infection in vitro. Finally, using a complete site-saturation mutagenesis methodology focused on the CDR (complementarity determining regions), a single point mutation (51E7) was identified that improved the 80% plaque reduction neutralization of the virus by greater than 8-fold. These discovery and evolution strategies can be applied to any emerging pathogen or toxin where a causative agent is known.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Viral / genetics
  • Antibodies, Viral / isolation & purification*
  • Antibodies, Viral / therapeutic use
  • Antibody Specificity / immunology
  • Chlorocebus aethiops
  • Communicable Diseases, Emerging / prevention & control
  • Communicable Diseases, Emerging / therapy*
  • Directed Molecular Evolution / methods*
  • Drug Discovery
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Neutralization Tests
  • Point Mutation / immunology
  • Severe Acute Respiratory Syndrome / prevention & control
  • Severe Acute Respiratory Syndrome / therapy*
  • Severe acute respiratory syndrome-related coronavirus / immunology*
  • Vero Cells

Substances

  • Antibodies, Viral