Sensitive Time-Gated Immunoluminescence Detection of Prostate Cancer Cells Using a TEGylated Europium Ligand

Anal Chem. 2016 Oct 4;88(19):9564-9571. doi: 10.1021/acs.analchem.6b02191. Epub 2016 Sep 15.

Abstract

We describe the application of a synthetically developed tetradentate β-diketonate-europium chelate with high quantum yield (39%), for sensitive immunodetection of prostate cancer cells (DU145). MIL38 antibody, a mouse monoclonal antibody against Glypican 1, conjugated directly to the chelate via lysine residues, resulted in soluble (hydrophilic) and stable immunoconjugates. Indirect labeling of the antibody by a europium chelated secondary polyclonal antibody and a streptavidin/biotin pair was also performed. All of these bright luminescent conjugates were used to stain DU145 cells, a prostate cancer cell line, using time gated luminescence microscopy for imaging, and their performances were compared to conventional FITC labeling. For all prepared conjugates, the europium chelate in conjunction with a gated autosynchronous luminescence detector (GALD) completely suppressed the cellular autofluorescence background to allow capture of vivid, high contrast images of immune-stained cancer cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Cell Line, Tumor
  • Coordination Complexes / chemical synthesis
  • Coordination Complexes / pharmacology*
  • Europium / chemistry*
  • Glypicans / immunology
  • Humans
  • Immunoconjugates / chemistry
  • Immunoconjugates / immunology
  • Immunoconjugates / pharmacology*
  • Immunologic Techniques / methods*
  • Ligands
  • Luminescence
  • Luminescent Agents / chemical synthesis
  • Luminescent Agents / pharmacology*
  • Male
  • Prostatic Neoplasms / diagnosis*

Substances

  • Antibodies, Monoclonal
  • Coordination Complexes
  • GPC1 protein, human
  • Glypicans
  • Immunoconjugates
  • Ligands
  • Luminescent Agents
  • Europium