Isolation and purification of vascular endothelial growth factor (VEGF) from ascitic fluid of ovarian cancer patients.
Vascular Endothelial Growth Factor (VEGF) or Vascular Permeability Factor (VPF) is an angiogenic cytokine expressed by many human and animal tumors. Because of the importance of VEGF in animal tumors, we purified VEGF/VPF from ascitic fluid of ovarian cancer patients with heparin sepharose column. The purified protein gave protein bands of 37 and 26 kD, respectively in 12% SDS PAGE. The specificity of the purified protein was determined with dot blot, trans-immunoblot and ELISA using polyclonal goat anti-VEGF antibody (Santa Cruz Biotechnology). The vasodilatatory effect of the purified protein was confirmed by a vascular permeability assay on mouse. A polyclonal mouse antibody was raised against the purified protein, which recognized the same protein by ELISA, transimmunoblot and dot-blot analysis. It has been also found that the raised polyclonal antibody in mouse- and the commercial VEGF polyclonal antibody (Santa Cruz Biotechnology) both inhibited in vitrocell proliferation of human MCF-7 cell line. This study shows for the first time an effort to purify VEGF from human source.
Ghosh
Sonali
S
Department of Metabolic Regulation, Chittaranjan National Cancer Institute, Kolkata, India.
Maity
Putul
P
D000328:Adult; D000818:Animals; D001202:Ascitic Fluid / Q000737:chemistry*; D015153:Blotting, Western; D049109:Cell Proliferation; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000276:immunology; D047468:Immunoprecipitation; D051379:Mice; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000737:chemistry* / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A / Q000302:isolation & purification*; D014664:Vasodilation
D007074:Immunoglobulin G; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A