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Identification of prostate specific membrane antigen (PSMA) as the target of monoclonal antibody 107-1A4 by proteinchip; array, surface-enhanced laser desorption/ionization (SELDI) technology.

Abstract
Recently we described the generation of the prostate tissue-specific monoclonal antibody (MAb) 107-1A4, its expression pattern and preliminary targeting of human prostate cancer xenografts. In this report we demonstrate that the target antigen for MAb 107-1A4 is prostate-specific membrane antigen (PSMA) using immunoaffinity absorption followed by SDS-PAGE and mass spectrometric analysis of peptides produced by in-gel tryptic digestion. The identity of the antigen has been confirmed by Western blots using MAbs of known specificity. MAb 107-1A4 is not reactive on Western blots. The conformational epitope for 107-1A4 is on the extracellular domain of PSMA. In competition studies, the binding of MAb 107-1A4 to LNCaP cells is inhibited by itself but not by any other of several other anti-PSMA MAbs, suggesting that the epitope may be unique. These results suggest that 107-1A4 is reactive to a conformational epitope in the external domain of PSMA that is unique among the panel of anti-PSMA MAbs in this study. Furthermore this work demonstrates the ability of mass spectroscopy to elucidate antibody-ligand interaction.
AuthorsS Wang, D L Diamond, G M Hass, R Sokoloff, R L Vessella
JournalInternational journal of cancer (Int J Cancer) Vol. 92 Issue 6 Pg. 871-6 (Jun 15 2001) ISSN: 0020-7136 [Print] United States
PMID11351309 (Publication Type: Journal Article)
CopyrightCopyright 2001 Wiley-Liss, Inc.
Chemical References
  • 107-1A4 monoclonal antibody
  • Antibodies, Monoclonal
  • Antigens, Surface
  • Epitopes
  • Ligands
  • Carboxypeptidases
  • FOLH1 protein, human
  • Glutamate Carboxypeptidase II
  • Trypsin
Topics
  • Antibodies, Monoclonal (analysis, chemistry)
  • Antigens, Surface
  • Binding, Competitive
  • Blotting, Western
  • Carboxypeptidases (chemistry)
  • Electrophoresis, Agar Gel
  • Electrophoresis, Polyacrylamide Gel
  • Epitopes
  • Glutamate Carboxypeptidase II
  • Humans
  • Ligands
  • Mass Spectrometry (methods)
  • Oligonucleotide Array Sequence Analysis
  • Precipitin Tests
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Tertiary
  • Trypsin (metabolism)
  • Tumor Cells, Cultured

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