Development of a lanthanide-based assay for detection of receptor-ligand interactions at the δ-opioid receptor

Heather L. Handl, Josef Vagner, Henry I. Yamamura, Victor J. Hruby, Robert J. Gillies

Research output: Contribution to journalArticle

39 Scopus citations

Abstract

A lanthanide-based assay for ligand-receptor interactions provides an attractive alternative to the traditional radiolabeled determinations in terms of sensitivity, throughput, and biohazards. We designed and tested five peptide ligands for the δ-opioid receptor that were modified with a europium (Eu)-containing chelate. These labeled ligands were tested for their binding affinities and compared with the unlabeled parental ligands. The Eu-diethylenetriaminepentaacetic acid (DTPA)-[d-Pen2,l-Cys 5] enkephalin (DPLCE) ligand bound to Chinese hamster ovary (CHO) cells overexpressing the human δ-opioid receptor with affinity similar to the unlabeled ligand. This ligand was used in competitive binding assays with results comparable to those obtained using the traditional radiolabeled binding assays. These lanthanide-based assays provide superior results with higher throughput and eliminate the need for radioactive waste disposal; hence, they are appropriate for high-throughput screening of ligand libraries.

Original languageEnglish (US)
Pages (from-to)299-307
Number of pages9
JournalAnalytical Biochemistry
Volume343
Issue number2
DOIs
StatePublished - Aug 15 2005

Keywords

  • Binding assay
  • DELFIA
  • DTPA
  • Lanthanide
  • Time-resolved fluorescence
  • δ-Opioid receptor

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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