Gender differentiation of bovine sperm cells
First Claim
1. A method for gender differentiation of bovine spermatozoa wherein monoclonal antibodies against male specific antigen, 17.18 kDa protein, associated with the classical pathway of the complement system are used after the inactivation of the alternative complement pathway, comprising:
- a. incubating heated guinea pig serum with sperm cells and monoclonal antibodies, said monoclonal antibodies being specific enough to recognize only male specific antigens, and b. selectively rupturing the male sperm cells, leaving a majority of the live sperm cells as female.
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Abstract
A method to make an immuno-differentiation of bovine sperm cells using monoclonal antibodies against male specific antigen, 17.18 kDa protein, associated to proteins of the classic pathway complement system is described. Monoclonal antibodies directed towards a gender-specific protein associated to the complement, provoke cellular rupture. For this methodology to be useful, an elimination of the alternative pathway of the complement is necessary. The sperm cells are then put in contact with a monoclonal antibody specifically for the gender-marking protein. Thus, the classic pathway complement system causes selected rupture of the marked sperm cells. The sperm cells of the treated semen are then ready to be used in artificial insemination, cryo-preservation or “in vitro” fertilization.
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Citations
14 Claims
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1. A method for gender differentiation of bovine spermatozoa wherein monoclonal antibodies against male specific antigen, 17.18 kDa protein, associated with the classical pathway of the complement system are used after the inactivation of the alternative complement pathway, comprising:
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a. incubating heated guinea pig serum with sperm cells and monoclonal antibodies, said monoclonal antibodies being specific enough to recognize only male specific antigens, and b. selectively rupturing the male sperm cells, leaving a majority of the live sperm cells as female. - View Dependent Claims (2, 3, 4, 5, 6)
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7. A method for gender differentiation of bovine spermatozoa wherein monoclonal antibodies against male specific antigen, 17.18 kDa protein, associated with the classical pathway of the complement system are used after the inactivation of the alternative complement pathway, using the following steps:
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a. Inactivating the alternative pathway of the complement system by heating guinea pig serum and ascitic fluid at 52.2°
C.+0.3 for 30 minutes or purified monoclonal antibodies; and
b. Incubating the sperm cells with monoclonal antibodies, originated from one or more different clones (hybridomas), for 45 minutes at 37°
C.; and
c. Washing the sperm cells to remove excess of antibodies, and then adding the inactivated guinea pig serum and seminal plasma to the sperm cell and then incubating for 1 hour at 37°
C. - View Dependent Claims (8, 9, 10, 11, 12, 13, 14)
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Specification