Use of specific monoclonal and polyclonal antibodies to define distinct antigens of the porcine zonae pellucidae

• Published in: Biology of reproduction Vol. 36; no. 5; pp. 1275 - 1287
• Main Authors: Timmons, T.M, Maresh, G.A, Bundman, D.S, Dunbar, B.S
• Format: Journal Article
• Language: English
• Published: Madison, WI Society for the Study of Reproduction 01-06-1987
• Subjects: Animals; Antibodies, Monoclonal - immunology; ANTICORPS MONOCLONAL; ANTICUERPOS MONOCLONALES; ANTIGENE; ANTIGENOS; ANTIGENS; Antigens - isolation & purification; Biological and medical sciences; CERDO; Egg Proteins; Electrophoresis, Polyacrylamide Gel; Epitopes - isolation & purification; Female; Fundamental and applied biological sciences. Psychology; GLICOPROTEINAS; GLYCOPROTEINE; GLYCOPROTEINS; Glycoproteins - immunology; Glycoproteins - isolation & purification; Hormone metabolism and regulation; Isoelectric Focusing; Mammalian female genital system; Membrane Glycoproteins; MONOCLONAL ANTIBODIES; OVA; OVULE; OVULO; Ovum - immunology; PORCIN; Rabbits; Receptors, Cell Surface; SWINE; Vertebrates: reproduction; Zona Pellucida - immunology; Zona Pellucida Glycoproteins; Glycoproteins; Monoclonal antibody; Zona pellucida; Pig; Antigen; Ovary; Vertebrata; Mammalia; Investigation method; Immunological investigation; Female genital system; Artiodactyla; Ungulata; Oocyte
• ISSN: 0006-3363; 1529-7268
• DOI: 10.1095/biolreprod36.5.1275

Specific monoclonal and polyclonal antibodies to solubilized porcine and rabbit zonae pellucidae (ZP) and to purified ZP glycoprotein
components have been used to define distinct ZP antigens. These studies demonstrate that the individual ZP glycoproteins contain
both unique and shared determinants. One monoclonal antibody (R5) has been used to demonstrate that the major porcine ZP glycoprotein,
which has multiple charge species ranging in molecular weight from 42,000 to 120,000, is composed of two distinct polypeptide
antigens unique to this glycoprotein class. These distinct antigens can be differentiated by immunoblotting after high-resolution
two-dimensional polyacrylamide gel electrophoretic separation of trypsin-treated or deglycosylated glycoproteins. The two
polypeptides also differ in their staining properties with the silver-based color stain and in their susceptibility to proteolysis.
A second monoclonal antibody (PSI) has been used to define a determinant shared by all three major porcine ZP glycoprotein
classes. This determinant appears to involve either a carbohydrate moiety or some other molecular feature related to post-translational
modification, since the antibody recognizes only the acidic species of each glycoprotein class, and does not recognize the
deglycosylated forms of the proteins. This work demonstrates that there are both unique and shared antigenic determinants
present in the individual components of the ZP, but that the immunodominant determinants appear to be unique to each glycoprotein.