A monoclonal anti-idiotypic antibody to μ and δ opioid receptors

Carmine J. Coscia, Maria Szücs, Jacob Barg, Mariana M. Belcheva, Wojciech T. Bem, Kamran Khoobehi, Tracy A. Donnigan, Regina Juszczak, Robert J. McHale, Michael R. Hanley, Eric A. Barnard

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Abstract

A mouse monoclonal, anti-idiotypic, anti-opioid receptor antibody (Ab2-AOR) has been generated from monoclonal anti-morphine antibodies (Ab1). Hybridoma culture supernatants were screened by a solid phase radioimmunoassay (RIA), based on their competition with radiolabelled morphine for Ab1. One of the Ab2s that gave a positive RIA also competed at rat brain opioid receptors with tritiated opioid ligands dihydromorphine (DHM), naloxone, etorphine, Tyr-d-Ala-Gly-Phe-d-Leu (DADLE), Tyr-d-Ala-Gly-NMe-Phe-Gly-ol (DAMGE) and Tyr-d-Pen-Gly-Phe-d-Pen (DPDPE). SDS-PAGE revealed Ab2-AOR to be highly purified after successive affinity and protein A-Sepharose chromatography. Ab2-AOR at concentrations of 10-100 nM competed with both μ- and δ-selective specific ligands for brain opioid receptors. Less than 13 μg/ml Ab2-AOR completely inhibited specific opioid radioligand binding to both soluble and membrane-bound opioid receptors. To demonstrate its anti-δ receptor activity further, a double-antibody ELISA procedure was developed that is based on the binding of Ab2-AOR to immobilized NG 108-15 cells (which contain only δ opioid receptors). Dose-dependent, opioid peptide- and opiate alkaloid-competitive binding of Ab2-AOR-containing ascites fluid to NG 108-15 cells was observed. A μ opioid agonist effect was demonstrated for Ab2-AOR, in that it decreased by 70% [3H]thymidine incorporation into DNA of fetal brain cell aggregates. This agonist-like action of Ab2-AOR was blocked by naltrexone. The antibody bound specifically to brain tissue sections and the presence of diprenorphine blocked this interaction. Hence, an Ab2 with μ and δ specificity has been characterized.

Original languageEnglish
Pages (from-to)299-306
Number of pages8
JournalMolecular Brain Research
Volume9
Issue number4
DOIs
Publication statusPublished - Mar 1991

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Keywords

  • ELISA
  • Hybridoma
  • Monoclonal anti-idiotypic antibody
  • Opioid peptide
  • Opioid receptor
  • Radioimmunoassay
  • Tissue print

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

Cite this

Coscia, C. J., Szücs, M., Barg, J., Belcheva, M. M., Bem, W. T., Khoobehi, K., Donnigan, T. A., Juszczak, R., McHale, R. J., Hanley, M. R., & Barnard, E. A. (1991). A monoclonal anti-idiotypic antibody to μ and δ opioid receptors. Molecular Brain Research, 9(4), 299-306. https://doi.org/10.1016/0169-328X(91)90076-A