In vivo far-red luminescence imaging of a biomarker based on BRET from Cypridina bioluminescence to an organic dye

Proc Natl Acad Sci U S A. 2009 Sep 15;106(37):15599-603. doi: 10.1073/pnas.0908594106. Epub 2009 Sep 8.

Abstract

We aimed to develop a far-red luminescence imaging technology for visualization of disease specific antigens on cell surfaces in a living body. First, we conjugated a far-red fluorescent indocyanine derivative to biotinylated Cypridina luciferase. This conjugate produced a bimodal spectrum that has long-wavelength bioluminescence emission in the far-red region as a result of bioluminescence resonance energy transfer. To generate a far-red luminescent probe with targeting and imaging capabilities of tumors, we then linked this conjugate to an anti-human Dlk-1 monoclonal antibody via the biotin-avidin interaction. This far-red luminescent probe enabled us to obtain high-resolution microscopic images of live, Dlk-1-expressing Huh-7 cells without an external light source, and to monitor the accumulation of this probe in tumor-bearing mice. Thus this far-red luminescent probe is a convenient analytical tool for the evaluations of monoclonal antibody localization in a living body.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • Biomarkers / metabolism
  • Biotin
  • Calcium-Binding Proteins
  • Cell Line, Tumor
  • Crustacea / enzymology
  • Energy Transfer*
  • Humans
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intercellular Signaling Peptides and Proteins / immunology
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Luciferases*
  • Luminescence
  • Luminescent Agents
  • Luminescent Proteins / genetics
  • Luminescent Proteins / immunology
  • Male
  • Membrane Proteins / genetics
  • Membrane Proteins / immunology
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Nude
  • Molecular Sequence Data
  • Neoplasms, Experimental / metabolism
  • Spectrophotometry

Substances

  • Antibodies, Monoclonal
  • Biomarkers
  • Calcium-Binding Proteins
  • DLK1 protein, human
  • Intercellular Signaling Peptides and Proteins
  • Luminescent Agents
  • Luminescent Proteins
  • Membrane Proteins
  • luciferase, cypridina
  • Biotin
  • Luciferases