Bispecific antibodies with natural architecture produced by co-culture of bacteria expressing two distinct half-antibodies

Nat Biotechnol. 2013 Aug;31(8):753-8. doi: 10.1038/nbt.2621. Epub 2013 Jul 7.


By enabling the simultaneous engagement of two distinct targets, bispecific antibodies broaden the potential utility of antibody-based therapies. However, bispecific-antibody design and production remain challenging, owing to the need to incorporate two distinct heavy and light chain pairs while maintaining natural nonimmunogenic antibody architecture. Here we present a bispecific-antibody production strategy that relies on co-culture of two bacterial strains, each expressing a half-antibody. Using this approach, we produce 28 unique bispecific antibodies. A bispecific antibody against the receptor tyrosine kinases MET and EGFR binds both targets monovalently, inhibits their signaling, and suppresses MET and EGFR-driven cell and tumor growth. Our strategy allows rapid generation of bispecific antibodies from any two existing antibodies and yields milligram to gram quantities of bispecific antibodies sufficient for a wide range of discovery and preclinical applications.

MeSH terms

  • Antibodies, Bispecific / biosynthesis*
  • Antibodies, Bispecific / immunology
  • Antibody Specificity
  • Bacteria / immunology
  • Bacteria / metabolism
  • Cell Line, Tumor
  • Coculture Techniques*
  • ErbB Receptors / genetics
  • ErbB Receptors / immunology*
  • Gene Expression Regulation, Bacterial / immunology
  • Humans
  • Immunoglobulin G / genetics
  • Immunoglobulin G / immunology
  • Neoplasms / immunology
  • Neoplasms / pathology
  • Neoplasms / therapy*
  • Protein Engineering
  • Proto-Oncogene Proteins c-met / genetics
  • Proto-Oncogene Proteins c-met / immunology*


  • Antibodies, Bispecific
  • Immunoglobulin G
  • EGFR protein, human
  • ErbB Receptors
  • MET protein, human
  • Proto-Oncogene Proteins c-met