Expanding the utility of beta-galactosidase complementation: piece by piece

Mol Pharm. 2010 Feb 1;7(1):60-74. doi: 10.1021/mp900188e.


The ability to image and quantify multiple biomarkers in disease necessitates the development of split reporter fragment platforms. We have divided the beta-galactosidase enzyme into unique, independent polypeptides that are able to reassemble and complement enzymatic activity in bacteria and in mammalian cells. We created two sets of complementing pairs that individually have no enzymatic activity. However, when brought into close geometric proximity, the complementing pairs associated resulting in detectable enzymatic activity. We then constructed a stable ligand complex composed of reporter fragment, linker, and targeting moiety. The targeting moiety, in this case a ligand, allowed cell surface receptor targeting in vitro. Further, we were able to simultaneously visualize two cell surface receptors implicated in cancer development, epidermal growth factor receptor and transferrin receptor, using complementing pairs of the ligand-reporter fragment complex.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers, Tumor / chemistry*
  • Biomarkers, Tumor / genetics*
  • Biomarkers, Tumor / metabolism
  • Cell Line, Tumor
  • ErbB Receptors / chemistry
  • ErbB Receptors / genetics
  • ErbB Receptors / metabolism
  • Escherichia coli K12 / genetics
  • Escherichia coli K12 / metabolism
  • Genes, Reporter
  • Genetic Complementation Test
  • Glioma / genetics
  • Glioma / metabolism
  • Humans
  • In Vitro Techniques
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Rats
  • Receptors, Transferrin / chemistry
  • Receptors, Transferrin / genetics
  • Receptors, Transferrin / metabolism
  • Transfection
  • beta-Galactosidase / chemistry*
  • beta-Galactosidase / genetics*
  • beta-Galactosidase / metabolism


  • Biomarkers, Tumor
  • Peptide Fragments
  • Receptors, Transferrin
  • ErbB Receptors
  • beta-Galactosidase