Site-specific immobilization of antibodies by their oligosaccharide moieties to new hydrazide derivatized solid supports.
This report describes a new method for immobilization of antibodies to solid supports. Antibodies are bound to the solid supports by covalent bonds between aldehydes generated on the carbohydrate side chains of the antibody and hydrazide groups on the solid support. The hydrazone bonds that are formed are stable at least from pH 2-10, permitting the acid elution of antigens from the affinity column. Over 25 mg of affinity-purified rabbit IgG binds per ml of solid support, with most of the bound antibodies retaining biological activity. Advantages of this new affinity support over existing technology are discussed along with procedures for the preparation and use of affinity columns containing monoclonal or polyclonal antibodies.
Hoffman
W L
WL
Department of Internal Medicine, Southwestern Medical School, Dallas, TX 75235.
O'Shannessy
D J
DJ
D000818:Animals; D000915:Antibody Affinity; D002236:Carbohydrate Conformation; D002846:Chromatography, Affinity / Q000379:methods; D000939:Epitopes / Q000276:immunology*; D006041:Goats; D006801:Humans; D006834:Hydrazines; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G; D051379:Mice; D009844:Oligosaccharides; D011817:Rabbits; D012685:Sepharose / Q000031:analogs & derivatives
D000939:Epitopes; D006834:Hydrazines; D007074:Immunoglobulin G; D009844:Oligosaccharides; D012685:Sepharose