A two-in-one antibody engineered from a humanized interleukin 4 antibody through mutation in heavy chain complementarity-determining regions

MAbs. May-Jun 2014;6(3):622-7. doi: 10.4161/mabs.28483. Epub 2014 Mar 11.

Abstract

A mono-specific antibody may recruit a second antigen binding specificity, thus converting to a dual-specific Two-in-One antibody through mutation at the light chain complementarity-determining regions (CDRs). It is, however, unknown whether mutation at the heavy chain CDRs may evolve such dual specificity. Herein, we examined the CDRs of a humanized interleukin 4 (IL4) antibody using alanine scanning and structural modeling, designed libraries of mutants in regions that tolerate mutation, and isolated dual specific antibodies harboring mutation at the heavy chain CDRs only. We then affinity improved an IL4/IL5 dual specific antibody to variants with dissociation constants in the low nanomolar range for both antigens. The results demonstrate the full capacity of antibodies to evolve dual binding specificity.

Keywords: Dual Action Fab; IL5; bispecific antibody; dual specific antibody; phage display.

MeSH terms

  • Animals
  • Antibodies, Bispecific / chemistry
  • Antibodies, Bispecific / genetics*
  • Antibodies, Monoclonal, Humanized / chemistry
  • Antibodies, Monoclonal, Humanized / genetics*
  • Binding Sites, Antibody / genetics
  • Complementarity Determining Regions / chemistry
  • Complementarity Determining Regions / genetics
  • Humans
  • Immunoglobulin Heavy Chains / chemistry
  • Immunoglobulin Heavy Chains / genetics
  • Interleukin-4 / immunology*
  • Mice
  • Models, Molecular
  • Mutagenesis
  • Peptide Library
  • Protein Engineering

Substances

  • Antibodies, Bispecific
  • Antibodies, Monoclonal, Humanized
  • Complementarity Determining Regions
  • IL4 protein, human
  • Immunoglobulin Heavy Chains
  • Peptide Library
  • Interleukin-4