Zona Pellucida Composition: Species Cross Reactivity and Contraceptive Potential of Antiserum to a Purified Pig Zona Antigen (PPZA)

Abstract

Pavlok, 1975; Oikawa and Yanagimachi, 1975)

The polypeptide backbone of recombinant human zona pellucida glycoprotein-3 initiates acrosomal exocytosis in human spermatozoa in vitro

“…The reagents used for the following procedures were of the highest standard commercially available and were obtained from Sigma (Poole, Dorset, U.K.), Fisher (Loughborough, Leics., U.K.) and BDH (Lutterworth, Leics., U.K. been shown to cross-react strongly with both intact human zonae pellucidae [13] and purified recombinant human ZP3 [14]. The Hamilton-Thorne motility analyser (HTM model 2030 ; Hamilton-Thorn Research Inc, Danvers, MA, U.S.A.) was used to assess spermatozoa motility before and after the following experimental procedures (see Clements et al [15] for HTM set up and quality control measures).…”

Milestones in contraceptive vaccines development and hurdles in their application

“…For example, antibodies generated against porcine ZP3 showed immunological cross-reactivity with human ZP. 93 Active immunization of females using various animal models with either the crude porcine zonae pellucidae or the purified native porcine zona proteins led to the curtailment of fertility. [94][95][96][97][98][99] Observed infertility was invariably associated with ovarian dystrophy accompanied by follicular attresia.…”

Immunoelectrophoretic analysis of the porcine zona pellucida

“…Four different preparations of solubilized porcine zonae pellucidae were used as antigens in electrophoretic studies: (1) SIZP (heat-solubilized isolated zonae pellucidae), (2) PPZA (purified pig zonae antigen), (3) and (4) Zonae encased pig eggs and/or isolated zonae were obtained essentially as described by Dunbar & Raynor (1980) and as modified by Subramanian et al (1981) and Sacco et al (1981). Briefly, quantities of 500 frozen-thawed pig ovaries, obtained from a local slaughterhouse, were homogenized in a commercial meat grinder in 50 mM-Tris-HCl buffer, pH 80, and isolated oocytes were collected by sieving the ovarian homogenate through nylon monofilament screens (Tetko, Inc.) Lowry, Rosebrough, Farr & Randall (1951) using bovine serum albumin as standard.…”

“…Pooled fractions of zona antigen-active material from this column were concentrated by ultrafiltration and are referred to as PPZA. Its purity was assessed by 2-D PAGE and gel photographs of a previous preparation of this isolated pig zona glycoprotein component have been published Sacco et al, 1981).…”

“…Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) of pig zonae has revealed the presence of three major components Sacco, Yurewicz, Subramanian & DeMayo, 1981 ; Dunbar, Liu & Sammons, 1981 ;Hedrick & Wardrip, 1981) with average molecular weights (Mr) of 55 000-60 000,65 000-70 000 and 90 000-92 000. Each of these components resolves into a family of charge isomers in the isoelectric focussing dimension with molecular weights of individual members increasing towards the acidic end of the isoelectric focussing gel in a manner which is highly characteristic of glycoproteins.…”