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Biology of Reproduction, Vol 24, 933-943, Copyright © 1981 by Society for the Study of Reproduction

Specific Radioimmunoassay for the Detection of a Purified Porcine Zona Pellucida Antigen (PPZA)

M. G. SUBRAMANIAN 1, E. C. YUREWICZ 1, , and A. G. SACCO 1

1 Department of Gynecology and Obstetrics, Wayne State University School of Medicine, The C. S. Mott Center for Human Growth and Development, Detroit, Michigan 48201


A radioimmunoassay (RIA) utilizing a biochemically purified porcine zona pellucida antigen (purified porcine zona antigen, PPZA) and rabbit antibodies raised against PPZA is described and characterized. PPZA was used for 125I labeling and as reference material in the RIA. Anti-PPZA activity was titered in serial dilutions of sera collected at weekly intervals for 11 weeks following the initial series of immunization injections and for 5 weeks following a single booster injection. This antiserum was of high titer, high affinity, and devoid of significant reactivity against the porcine tissues tested; however, a measurable but low level of cross reactivity was observed when tested against pig serum and pig follicular fluid. In response to the initial immunization series, peak titer was obtained in 6 weeks at which time 30% of [125I]-PPZA was precipitated by antibody diluted to 1:400,000. There was no further increase in antibody titer following the booster immunization. Details of PPZA iodination, stability of [125I]-PPZA, RIA methodology, and assay validation for sensitivity, precision, accuracy, specificity, and other parameters are described.

Note:
ACKNOWLEDGMENTS The technical assistance of Jacqueline Patt and Larry Rothstein is gratefully acknowledged. This research was supported by Grant 5 RO1 HD10189 from NICHD, NIH.

Submitted on October 21, 1980
Accepted on January 16, 1981




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[Abstract] [Full Text] [PDF]


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