Chemotactic factors for human spermatozoa and their use in human assisted fertilization
AssigneeYeda Research and Development Co. Ltd.
InventorMichael Eisenbach
Life of this patent
Estimating the term…
Abstract
The invention relates to chemotactic factors for human spermatozoa that are purifiable from human follicular fluid. The factors are of peptidic and hydrophilic nature and have, one, a molecular size of about 13 kDa and, the other, an apparent molecular size smaller than 1.3 kDa, as determined by high pressure gel filtration. They are for use in procedures related to human fertilization, such as in various types of assisted fertilization, particularly artificial insemination, in vitro fertilization, micromanipulation and direct microinjection of sperm into oocytes.
Claims21
A purified chemotactic factor for human spermatozoa purifiable from human follicular fluid, said factor being of peptidic and of hydrophilic nature characterized by:
- (i) it is selected from a group consisting of a factor having a molecular size of about 13 kDa and a factor having an apparent molecular size smaller than 1.3 kDa, the molecular sizes being determined by high pressure gel filtration;
- (ii) it causes human sperm chemotaxis and chemokinesis and, ultimately, hyperactivation-like motility;
- (iii) its specific sperm chemotactic activity is concentration dependent;
- (iv) when purified from human follicular fluid or from a 90% acetone supernatant thereof by high pressure gel filtration as in (i) above, the 13 kDa chemotactic factor is eluted at 17.0-17.5 min and the <1.3 kDa chemotactic factor is eluted at 25.0-25.5 min, the specific sperm chemotactic activity (per protein content) of each factor being at least 1,000 fold higher than that of the follicular fluid from which said factor was Purified;
- and
- (v) when purified by reversed-phase HPLC from a supernatant of human follicular fluid precipitated with 90% acetone or from the active fractions eluted from the column described in (i) above, the sperm chemotactic activity of the 13 kDa factor is eluted at 20% acetonitrile, and the sperm chemotactic activity of the <1.3 kDa factor is eluted at 0% acetonitrile, the specific chemotactic activity (per protein content) of each factor being at least 10,000 fold higher than that of the follicular fluid from which said factor was purified.
Dependencies are inferred from the claim text.