Ovarian cancer-associated antibodies recovered from ascites: their use for the isolation of ovarian cancer-associated antigen to produce monoclonal antibodies

Gynecol Oncol. 1990 Apr;37(1):24-8. doi: 10.1016/0090-8258(90)90301-z.

Abstract

Immune complexes (ICs) were recovered from the ascites of a patient with stage IV endometrioid ovarian cancer by sequential precipitation with 33% saturated ammonium sulfate and 2.5% polyethylene glycol 6000 (PEG 6000), followed by affinity chromatography on protein A-Sepharose CL-4B. The IgG-containing ICs were dissociated using 8 M urea, separated by ion-exchange chromatography on Sephadex QAE-50, and subsequently analyzed for purity by immunoelectrophoresis (IEP) and radial immunodiffusion (RID). Recovered antibody was tested for reactivity by immunohistologic techniques against paraffin-embedded tumor tissue and acetone-fixed cell suspensions of epithelial tumors. The antibody which demonstrated ovarian cancer-associated activity was absorbed with antigen extracts of breast, colon, and lung cancers as well as keratin to reduce cross-reactivity. The absorbed endometrioid ovarian cancer-associated antibody (OCAAb) was used to produce an immunoadsorbent column for the recovery of tumor-associated antigens. A mouse monoclonal antibody designated FEN-1 was produced using this antigen-containing fraction, and preliminary screening has demonstrated ovarian tumor-associated reactivity. The use of autologous ICs as reagents for preparing tumor antigen-rich immunogens may provide a valuable tool in the search for tumor-associated antigens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Neoplasm / isolation & purification*
  • Antigen-Antibody Complex / isolation & purification*
  • Antigens, Neoplasm / isolation & purification*
  • Ascitic Fluid / immunology*
  • Chromatography, Affinity
  • Cross Reactions
  • Female
  • Fluorescent Antibody Technique
  • Humans
  • Immunoenzyme Techniques
  • Neoplasm Staging
  • Ovarian Neoplasms / immunology*
  • Ovarian Neoplasms / pathology

Substances

  • Antibodies, Monoclonal
  • Antibodies, Neoplasm
  • Antigen-Antibody Complex
  • Antigens, Neoplasm
  • ovarian tumor associated antigen