A specific radioimmunoassay for evaluation of serum antibodies to zona pellucida antigens

Studies on the stability of 125I‐purified porcine zona antigen (PPZA) obtained by chloramine‐T glucose oxidase with lactoperoxidase and N‐bromosuccinimide lodination procedures

“…Over the years, immunocontraceptive studies involving the zona-pellucida were handicapped because of the lack of antigen(s) with sufficient purity. Following the description of an assay system utilizing a heterologous solubilized zona antigen for labeling and reference material [Palm et al, 19791, similar assay methodology was adopted by other investigators [Gerrity et al, 1981;Hulme et al, 19821. Further advancement was accomplished with the development of a specific and sensitive assay system which utilized a purified porcine zona antigen (PPZA) [Subramanian et al, 19811 and which was used in studies testing the contraceptive potential of PPZA [Sacco et al, 19811, the immune response to PPZA in a primate system [Sacco et al, 19831, and preliminary biochemical characterization [Yurewicz et al, 19831.…”

“…The application of a radioimmunoassay (RIA) system from this laboratory utilizing porcine zonae demonstrated for the first time the tissue specificity of zona pellucida antigen@) and the monospecificity of antibodies to isolated porcine zonae pellucidae [Palm et al, 19791. Subsequently, studies utilizing solubilized zonae material for radiolabeling were reported by others [Gerrity et al, 1981;Hulme et al, 19821. A RIA procedure utilizing a biochemically purified porcine zona antigen (PPZA: Mr = 58000) and rabbit antibodies raised against PPZA was described and characterized [Subramanian et al, 19811.…”

The Mammalian Egg Coat

“…Since earlier investigations (Yanagimachi et al 1976, Palm et al 1979Gerrity et al 1981) have clearly demonstrated the tissue specificity of zona antigens by the failure of zona antibodies to cross-react with other body tissues, the observed effects at the level of the ovary must be caused by a specific reaction of zona antibodies with zona components. However, the exact cellular target(s) of these zona antibodies and their mechanism of action in producing such disturbances in normal ovarian function can at present only be speculated.…”

“…The introduction of sensitive RIA techniques (Palm et al 1979) both for the detection of zona material, i.e., competitive binding assays Palm et al 1979), and for the detection and titration of zona antibodies, i.e., direct binding assays (Gerrity et al 1981;Sacco et al 1981 b), has resolved a major shortcoming of early immunocon.tracep· tive investigations involving the zona. The introduction of sensitive RIA techniques (Palm et al 1979) both for the detection of zona material, i.e., competitive binding assays Palm et al 1979), and for the detection and titration of zona antibodies, i.e., direct binding assays (Gerrity et al 1981;Sacco et al 1981 b), has resolved a major shortcoming of early immunocon.tracep· tive investigations involving the zona.…”

Use of a radioimmunoassay technique to measure the titer of anti‐cow‐zona antibodies in marmoset monkeys

“…The usual methods for assessing anti-zona antibodies, such as immunoprecipitation and immunofluorescence, are limited in scope and only quantitative to a limited extent, while in vitro fertilization, bioassay techniques [Aitken and Richardson, 1981bl are labor intensive and require careful standardization. More recently, however, radioimmunoassays have been developed for porcine zona antigens, which are specific for anti-zona antibodies raised in rabbits [Palm et al, 1979;Subramanian et al, 19811 and have been shown to be rapid, reproducible procedures for the detection of anti-zona antibodies in serum [Gerrity et al, 1981;Subramanian et al, 19811.…”