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Molecular Human Reproduction Vol. 2, NUMBER 12 pp. 911-919, 1996
© European Society of Human Reproduction and Embryology 1996

research-article

Biological activity of recombinant human ZP3 produced in vitro: potential for a sperm function test

A.J. Whitmarsh1,3, M.J. Woolnough2, H.D.M. Moore1,2, D.P. Hornby1 and C.L.R. Barratt2,4

1Krebs Institute, Department of Molecular Biology and Biotechnology, University of Sheffield PO Box 594, Firth Court, Western Bank, Sheffield, S10 2TN, UK 2University Department of Obstetrics and Gynaecology, Jessop Hospital for Women Sheffield S3 7RE, UK

To whom correspondence should be addressed at: 4To whom correspondence should be addressed

The human zona binding test is the most predictive test of sperm function yet the availability of human zona severely restricts its clinical use. The primary aim of this study was to use a commercially available in-vitro transcription and translation system to produce immobilized recombinant human ZP3 (rhuZP3) on agarose beads. The biological activity of this preparation was examined using sperm binding and the acrosome reaction. Significantly higher levels of sperm binding to rhuZP3 beads (n = 12, P <0.05) compared with controls were observed and there was a significant induction (n = 12, P <0.01) in the acrosome reaction after overnight incubation at 37°C in 5% CO2, in air. In conclusion, the in-vitro transcription and translation system can produce sufficient quantities of purified immobilized biologically active rhuZP3. These preliminary experiments will enable further refinements to be made so that a solid-phase sperm function test based on rhuZP3 coated beads is likely to be developed in the near future.

acrosome reaction/spermatozoa/zona/zona binding/ZP3

3Current address: Howard Hughes Medical Institute, Program of Molecular Medicine, University of Massachusetts Medical School, 373 Plantation Street, Worcester, MA 01605, USA


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