Characterization of Porcine Zona Pellucida Antigens by Immunoaffinity Chromatography and by High-Pressure Liquid Chromatography

Authors

  • JOHANNES DIETL,

    Corresponding author
    1. Department of Gynecology and Obstetrics, Christian-Albrechts University and Michaelis Midwifery School, Kiel, Federal Republic of Germany
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  • ADAM CZUPPON,

    1. Department of Gynecology and Obstetrics, Christian-Albrechts University and Michaelis Midwifery School, Kiel, Federal Republic of Germany
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  • LILO METTLER

    1. Department of Gynecology and Obstetrics, Christian-Albrechts University and Michaelis Midwifery School, Kiel, Federal Republic of Germany
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Universitats-Frauenklinik, Hegewischstrasse 4.2300 Kiel Federal Republic of Germany.

Abstract

ABSTRACT: 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/l are expressed on the surface in intact zonae.

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