The use of alginate ester films on solid supports in the preparation of water-insoluble immunoadsorbents for purification of antigens.
A novel method developed for the preparation of immunoadsorbents is based on the technique of coating potential column supports with a propylene glycol alginate ester film created by emulsion polymerisation. The cross-linking of the film with 1,2-diaminoethane in alkaline solution resulted in a water-permeable but water-insoluble film which was stable at high pH. Sheep anti-(human IgG) was subsequently coupled to the cyanogen-bromide-activated film and the resultant immunoadsorbents were used to purify IgG from whole human serum. The production of the film-coated supports, chemical coupling of antibody and the chromatographic characteristics of four alginate-film-coated supports are described. With the use of Sepharose--alginate-ester--sheep-anti-(human IgG) immunoadsorbent the overall yield for IgG from five serial adsorption-elution cycles was 94% of the adsorbed IgG. This immunoadsorbent proved to be of high binding capacity and allowed the elution of specific antigen in almost 100% yield and high purity with minimal non-specific interference from other serum proteins.
D000464:Alginates; D000818:Animals; D000936:Antigen-Antibody Complex; D000941:Antigens / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D007164:Immunosorbents; D012756:Sheep; D012995:Solubility; D014867:Water
D000464:Alginates; D000936:Antigen-Antibody Complex; D000941:Antigens; D007074:Immunoglobulin G; D007164:Immunosorbents; D014867:Water; C038550:propylene glycol alginate ester