Inhibition of macrophage migration by a factor from ascites fluids of ovarian cancer patients. III. Generation of monoclonal antibody specific for human MIF.
Migration inhibition activity from ascitic fluids of ovarian cancer patients (OC-MIF) was used to develop monoclonal antibodies. The OC-MIF was purified about 10,000 fold by affinity chromatography on L-fucose-Sepharose 6B. Spleen cells from AB/Jena mice immunized with purified OC-MIF were hybridized with P3X63 Ag 8.653 myeloma cells. Supernatants of the hybridoma cultures were screened by solid-phase binding assay, direct neutralizing assay and solid-phase RIA. Several clones of these hybridomas secreted antibodies into the culture medium, which neutralized the biological activity of OC-MIF at dilutions as high as 10(-4) relative to the initial culture medium. After expansion and cloning one clone was selected for ascitic antibody production. This monoclonal antibody coupled to Sepharose 4B adsorbed OC-MIF. Most of the adsorbed biological activity could be eluted with 0.1 M acetic acid.
Fahlbusch
B
B
Zentralinstitut für Mikrobiologie und experimentelle Therapie der Akademie der Wissenschaften der Deutschen Demokratischen Republik, Jena.
Schumann
I
I
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D000918:Antibody Specificity; D001202:Ascitic Fluid / Q000032:analysis*; D002846:Chromatography, Affinity / Q000379:methods; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D008263:Macrophage Migration-Inhibitory Factors / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D009500:Neutralization Tests; D010051:Ovarian Neoplasms / Q000032:analysis*
D000911:Antibodies, Monoclonal; D008263:Macrophage Migration-Inhibitory Factors