Methods and reagents for evaluating the binding of mammalian sperm

Patent metadata

Field	Value
Publication number	US-9594077-B2
Application number	US-201314412318-A
Country	US
Kind code	B2
Filing date	Jul 3, 2013
Priority date	Jul 3, 2012
Publication date	Mar 14, 2017
Grant date	Mar 14, 2017

Abstract

The present invention provides a new apparatus/system for determining fertility of mammalian sperm based on a specific sperm-binding test that mimics sperm binding to an egg. The inventive apparatus/system comprises surface-modified beads with a compound or conjugates containing a carbohydrate sequence associated with the mammalian zona-pellucida (ZP) glycoproteins.

First claim

What claimed is:

1. A method of evaluating the ability of non-human mammalian sperm to bind to a mammalian egg, the method comprising:
(a) combining a sample of non-human mammalian sperm, a media that is conducive to maintaining the physiological activity of the sperm, and at least one substrate that comprises bovine thyroglobulin on at least a portion of its surface, to allow for at least a portion of the sample of non-human mammalian sperm to bind to the substrate, wherein the binding of sperm to the substrate is indicative of the binding of sperm to a mammalian egg; and
(b) visualizing or quantitating the sperm bound to the substrate, thereby evaluating the ability of non-human mammalian sperm to bind to a mammalian egg.

2. The method of claim 1 further comprising removing at least a portion of the unbound sperm before visualizing or quantitating the sperm bound to the substrate.

3. The method of claim 1 wherein the substrate comprises a crosslinked agarose bead.

4. The method of claim 1 wherein the compound or conjugate is attached to the surface of a specimen slide, dish, or well of a microtiter plate.

5. The method of claim 1 wherein the compound or conjugate is fixed for use in flow cytometry.

6. The method of claim 1 , wherein the mammalian sperm is equine or porcine.

7. The method of claim 1 , wherein the mammalian sperm is porcine.

Assignees

• Univ Missouri

Inventors

• Clark Gary Francis
• Sutovsky Peter

Classifications

• G01N2400/38

  Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence, e.g. gluco- or galactomannans, Konjac gum, Locust bean gum or Guar gum · CPC title

• G01N2800/367

  Infertility, e.g. sperm disorder, ovulatory dysfunction · CPC title

• G01N33/5091 Primary

  for testing the pathological state of an organism · CPC title

• G01N33/50 Primary

  Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing (measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q) · CPC title

• G01N33/56966

  Animal cells · CPC title