Inhibition of macrophage migration by a factor from ascites fluids of ovarian cancer patients. II. Production and characterization of an anti-MIF antibody.
A macrophage migration inhibition factor (OC-MIF) has been isolated from ascites fluid of ovarian cancer patients by affinity chromatography on L-fucose-Sepharose 6B, and characterized biochemically. OC-MIF activity was purified approximately 10 000-fold as compared to the starting material. It exhibits molecular heterogeneity with respect to net charge and molecular weights. Compared to it, purified and radioiodinated OC-MIF is fairly homogeneous and contains a major protein component with a molecular mass of about 45 kD, and two isoelectric points of 3.0-4.0 and about 5.0. Rabbits were immunized with the highly purified MIF material and an antiserum was prepared and was used to prepare immunoadsorbent beads. Beads made with anti-OC-MIF antiserum, but not with rabbit control serum, could remove specifically OC-MIF activity and showed weak reactivity towards Con A induced MIF. Using a radioimmunoassay (RIA) anti-OC-MIF antiserum reacts with OC-MIF and also with Con A induced MIF. This antigenic relationship between conventional MIF and OC-MIF and common biochemical properties suggest that the two mediator substances are very similar and may, perhaps, be identical. Furthermore, the possibility to determine various MIF activities by means of RIA was investigated.
D000906:Antibodies / Q000302:isolation & purification*; D001201:Ascites; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003208:Concanavalin A / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007163:Immunosorbent Techniques; D007525:Isoelectric Focusing; D008263:Macrophage Migration-Inhibitory Factors / Q000276:immunology*; D008264:Macrophages / Q000187:drug effects; D010051:Ovarian Neoplasms / Q000032:analysis*; D011863:Radioimmunoassay
D000906:Antibodies; D008263:Macrophage Migration-Inhibitory Factors; D003208:Concanavalin A