Abstract: Reagents, methods, and kits are described that are useful for fertility testing and that in many cases provides much faster, convenient and rapid determination of male infertility. The reagents described include properly glycosylated human ZP3, ZP2, ZP1, glycosylated peptides thereof, fusion proteins such as green fluorescent protein—ZP3, non-covalent complexes of ZP2 with ZP3, fusion protein of ZP2–ZP3, solid phase materials such as agarose beads coated with binding agents such as ZP3, and other artificial zona. Methods are provided that, in many cases convert a complex biological event into a well defined biochemical binding event based on one or more of the reagents. Such methods are much easier to set up and monitor, allowing more convenient and inexpensive diagnostic testing for male fertility. The acrosome reaction is detected in other embodiments by virtue of quantitating one or more released substances.

Abstract: The present invention provides a method to determine sperm activity comprising the steps of: (a) contacting an appropriate concentration of human zona pellucida protein 3 with an appropriate amount of sperm under conditions permitting the formation of a complex between the human zona pellucida protein 3 and the sperm; and (b) determining the complex formed. The invention further provides a method to determine sperm activity comprising the steps of (a) contacting an appropriate concentration of human zona pellucida protein 3 with an appropriate amount of sperm under conditions permitting an acrosome reaction to occur; and (b) determining the extent of the acrosome reaction.

Abstract: The invention relates to methods for producing recombinant human zona pellucida protein (“rhZP3”) and glycosylated peptide having biological activity of binding to human spermatozoa. A human ovarian cell line is used to produce rhZP3 having a glycosylation pattern required for full biological activity. Methods of determining useful peptides with binding activity for human sperm and their syntheses, as well as using such peptides and proteins in therapeutics and diagnostics are discussed.

Abstract: The invention relates to methods for producing recombinant human zona pellucida protein (“rhZP3”) and glycosylated peptide having biological activity of binding to human spermatozoa. A human ovarian cell line is used to produce rhZP3 having a glycosylation pattern required for full biological activity. Methods of determining useful peptides with binding activity for human sperm and their syntheses, as well as using such peptides and proteins in therapeutics and diagnostics are discussed.

Abstract: The present invention provides a method to determine sperm activity comprising the steps of: (a) contacting an appropriate concentration of human zona pellucida protein 3 with an appropriate amount of sperm under conditions permitting the formation of a complex between the human zona pellucida protein 3 and the sperm; and (b) determining the complex form. The invention further provides a method to determine sperm activity comprising the steps of (a) contacting an appropriate concentration of human zona pellucida protein 3 with an appropriate amount of sperm under conditions permitting an acrosome reaction to occur; and (b) determining the extent of the acrosome reaction.

Abstract: An environmentally controlled incubator is described in which the enclosure of the incubator chamber strongly attenuates the transmission of light having wavelengths below about 500 nm for protecting biological materials within the chamber from toxic effects of short wavelength light. The incubator also includes sensors for determining the oxygen and carbon dioxide concentration within the chamber and means for adding carbon dioxide, nitrogen or oxygen to the ambient gas within the incubator ion order to maintain the desired levels of carbon dioxide and oxygen. A thermostatically controlled heater and a humidification chamber warm and humidify the gas as it is circulated through the humidification chamber under control of a fan. An airlock is provided to allow biological materials to be placed within or removed from the chamber.