Efficient purification of GP-1D8 glycoprotein from human breast carcinoma tissue by immunoaffinity chromatography

Prep Biochem Biotechnol. 2008;38(4):321-33. doi: 10.1080/10826060802325204.

Abstract

Anti-GP-1D8 monoclonal antibody was produced in our laboratory. Immunoaffinity purification of GP-1D8 glycoprotein from human breast carcinoma tissues, on column with the monospecific antibody, is developed. The procedure permits purification of GP-1D8 to a highly purified state. It appeared as a single band in sodium dodcyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular mass of GP-1D8 was determined to be 66 kDa by mass spectrometry. Its antigenicity was stable between 0 and 70 degrees C. It was breast cancer specific, as determined by immunohistochemistry and immunocytochemistry. The preparation of anti-GP-1D8 monoclonal antibody will facilitate further detection of, and functional study on, GP-1D8. The study also provides a simple, economical, and efficient method for affinity purification of target proteins from human or animal tissues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Breast Neoplasms / metabolism*
  • Cell Fusion
  • Chromatography, Affinity / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Glycoproteins / immunology*
  • Glycoproteins / isolation & purification*
  • Humans
  • Immunohistochemistry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Antibodies, Monoclonal
  • GP-1D8 glycoprotein, human
  • Glycoproteins