Rapid and efficient purification of mouse monoclonal antibodies from ascites fluid using high performance liquid chromatography.
Techniques for the rapid and efficient purification of mouse monoclonal antibodies from murine ascites are described that utilize anion exchange and gel permeation chromatography using high performance liquid chromatography (HPLC). Anion exchange chromatography was performed at neutral pH using a hydrophilic resin conjugated with a substituted amine (Mono Q column, Pharmacia Fine Chemicals). Various subclasses of mouse IgG monoclonals were assessed for their binding to this matrix, with all of the antibodies tested eluting at relatively low concentrations of sodium chloride. In some instances, a protein tentatively identified as transferrin was co-purified using this anion exchange procedure. However, this protein was easily removed from the IgG using gel permeation chromatography (Bio-Sil TSK-250, Bio-Rad), also performed at neutral pH.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001202:Ascitic Fluid / Q000276:immunology*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange; D006041:Goats; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000302:isolation & purification; D051379:Mice; D011817:Rabbits; D012756:Sheep
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G