Purification of IgM monoclonal antibody from murine ascitic fluid by a two-step column chromatography procedure.
Two IgM monoclonal antibodies (Mabs) were purified from murine ascitic fluid by a two-step procedure involving pseudoligand affinity chromatography on immobilized Cibacron Blue F3GA and gel filtration chromatography on ACA-22. The purified IgM from one ascites sample, H-1, was determined to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis while ascites containing E3.1 IgM Mab, was purified to greater than 95% IgM. The retention of biological activity was confirmed by immunofluorescence staining of peripheral blood lymphocytes.
D000178:Acrylamides; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001202:Ascitic Fluid / Q000032:analysis*; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D007075:Immunoglobulin M / Q000302:isolation & purification*; D051379:Mice; D012685:Sepharose; D014227:Triazines
D000178:Acrylamides; D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M; D014227:Triazines; C012842:Cibacron Blue F 3GA; C052622:ACA 22 gel; D012685:Sepharose