Purification of monoclonal antibodies on dextran-coated silica support grafted by thiophilic ligand.
Coated silica beads are promising supports for high-performance liquid chromatography (HPLC) of proteins; they combine the excellent mechanical properties of silica with minimal non-specific interactions with proteins in solution due to the presence of hydrophilic dextran polymers adsorbed at the silica surface. So, dextran-coated porous silica beads can be grafted with beta-mercaptoethanol by using divinylsulfone as coupling reagent to obtain new thiophilic supports usable in HPLC. The affinity of monoclonal IgG subclasses from mouse ascitic fluids for the active phases can be analysed. These dextran-coated silica supports grafted with thiophilic ligands allow a one-step purification of these antibodies. Moreover, the chromatographic separation of two subclasses, immunoglobulin G1 (IgG1) and IgG3, is observed and can be correlated to the high resolution of these new HPLC thiophilic supports.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000915:Antibody Affinity; D001201:Ascites / Q000276:immunology; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003911:Dextrans / Q000737:chemistry*; D007074:Immunoglobulin G / Q000302:isolation & purification*; D008623:Mercaptoethanol / Q000737:chemistry; D051379:Mice; D012822:Silicon Dioxide / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D003911:Dextrans; D007074:Immunoglobulin G; D008623:Mercaptoethanol; D012822:Silicon Dioxide