Efficient Purification of GP-1D8 Glycoprotein from Human Breast Carcinoma Tissue by Immunoaffinity Chromatography

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°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.

Keywords:

• Breast cancer
• Glycoprotein
• Immunoaffinity chromatography
• Purification

ACKNOWLEDGMENT

We thank Professor Yuxian Zhu (College of Life Sciences, Peking University) for constructive discussions and giving us some help for accomplishing this work. This work was supported by National Natural Science Foundation of China (No. 30571711) to Renqing Feng.