Engineering of a broad-specificity antibody: detection of eight fluoroquinolone antibiotics simultaneously

Anal Biochem. 2011 Feb 1;409(1):14-21. doi: 10.1016/j.ab.2010.09.041. Epub 2010 Nov 4.


Recombinant sarafloxacin-recognizing antibody was engineered with the use of novel fluoroquinolone (FQ) derivatives. A monoclonal FQ antibody, 6H7, was targeted to random mutagenesis to broaden the specificity of the antibody in development of a generic assay for FQ antibiotics. Engineering involved the synthesis of different small-sized FQ molecules to immobilize and detect the mutant antibodies. Selections with labeled FQs resulted in several mutant antibodies with increased affinity or wider specificity toward different FQs. The best characterized mutant antibody was capable of recognizing seven of eight targeted FQs below maximum residue limits set by the European Union. The results are promising in regard to the development of a multiresidue immunoassay for FQs based on a single antibody.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / analysis*
  • Anti-Bacterial Agents / chemical synthesis
  • Anti-Bacterial Agents / immunology
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / metabolism
  • Antibody Specificity
  • Biotinylation
  • Fluoroquinolones / analysis*
  • Fluoroquinolones / chemical synthesis
  • Fluoroquinolones / immunology
  • Immunoassay / methods*
  • Mutagenesis
  • Peptide Library
  • Protein Engineering*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Structure-Activity Relationship


  • Anti-Bacterial Agents
  • Antibodies, Monoclonal
  • Fluoroquinolones
  • Peptide Library
  • Recombinant Proteins