Binding characteristics of sperm with recombinant human zona pellucida glycoprotein-3 coated beads.
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Date
2009-07
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Abstract
Background & objectives: An inability or decreased ability of spermatozoa to bind to the zona pellucida
(ZP), an extracellular glycoproteinaceous matrix surrounding egg, is one of the plausible causes of
idiopathic infertility. It will be clinically useful to distinguish this condition from other causes of infertility.
An assay system, investigating binding of human sperm with ZP glycoprotein may prove useful in this
regard. We attempted to develop a simple assay system to analyse the binding of capacitated human
spermatozoa to human zona pellucida glycoprotein-3 (ZP3) using baculovirus-expressed recombinant
human ZP3 coated beads.
Methods: Recombinant baculovirus-expressed ZP3 was purified, labelled with biotin and coated on
streptavidin sepharose beads. An in vitro assay system was optimized to study binding of capacitated
human sperm to ZP3 coated beads.
Results: A higher percentage of baculovirus-expressed recombinant human ZP3 coated beads showed
significant (P<0.05) binding of capacitated human sperm as compared to beads coated with fetuin. An
inhibition in the binding of sperm to ZP3 coated beads was observed in presence of cold recombinant
human ZP3. Further, prior incubation of ZP3 coated beads with monoclonal antibodies (MAbs) against
ZP3 but not against ZP2 resulted in the decrease in number of sperm bound to bead.
Interpretation & conclusion: An in vitro assay system to study the binding of human sperm to ZP3-
primary sperm receptor was established, which may be useful to determine the functional competence
of spermatozoa.
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Keywords
Baculovirus-expression system, human zona pellucida glycoprotein-3, monoclonal antibody, spermatozoa, streptavidin sepharose beads
Citation
Bansal Pankaj, Gupta S K. Binding characteristics of sperm with recombinant human zona pellucida glycoprotein-3 coated beads. Indian Journal of Medical Research. 2009 Jul; 130(1): 37-43.