Efficient purification of GP-1D8 glycoprotein from human breast carcinoma tissue by immunoaffinity chromatography.
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.
Feng
Renqing
R
Department of Biochemistry, College of Life Sciences, Peking University, Beijing, P R China. rqfeng@pku.edu.cn
Guo
Zhenquan
Z
Li
Jinfeng
J
D000911:Antibodies, Monoclonal / Q000276:immunology; D001943:Breast Neoplasms / Q000378:metabolism*; D002459:Cell Fusion; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000276:immunology* / Q000302:isolation & purification*; D006801:Humans; D007150:Immunohistochemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000911:Antibodies, Monoclonal; C532947:GP-1D8 glycoprotein, human; D006023:Glycoproteins