Characterization of porcine zona pellucida antigens by immunoaffinity chromatography and by high-pressure liquid chromatography.
In the present work, 500 and 50,000 porcine zonae pellucidae were solubilized using Lithium-3,5-diiodosalicylate. The zona antigens were purified by immunoaffinity chromatography (IAC) on immobilized antizona immunoglobulin G (IgG). The antizona-IgG was raised by immunization of female rabbits with 500 heat-solubilized porcine zonae. Four antigens could be detected following IAC: ZP I/1 (Mr = 42,000), ZP II/1 (Mr = 67,000), ZP II/2 (Mr = 32,000), ZP III/1 (Mr = 17,000). In a parallel experiment, 50,000 zonae were solubilized in a similar manner and the mixture was analyzed by high-pressure liquid chromatography (HPLC) using a protein column. Altogether, 9 protein peaks that contained the antigens ZP I/1, ZP II/1, ZP II/2, and ZP III/1 could be detected following HPLC. The carbohydrate composition is characteristic for O-glycosidic-glycoproteins. ZP II/1 and ZP II/2 are probably in close association within the zona. Based on the reaction of the antigens with antibodies induced by intact and heat-solubilized zonae, it is postulated that only ZP I/1 and ZP II/1 are expressed on the surface in intact zonae.
D000818:Animals; D000941:Antigens / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D005260:Female; D007074:Immunoglobulin G; D008970:Molecular Weight; D010063:Ovum / Q000276:immunology*; D011506:Proteins / Q000276:immunology; D011817:Rabbits; D013552:Swine / Q000276:immunology; D015044:Zona Pellucida / Q000276:immunology*
D000941:Antigens; D007074:Immunoglobulin G; D011506:Proteins