Zona pellucida binding peptides, expression vectors, compositions, and methods for species-specific immunocontraception of animals

Abstract

Disclosed are methods, compositions, zona pellucida binding peptides and polypeptides, and expression vectors for use in species-specific immunocontraception of animals. The disclosed compositions may include immunogenic compositions or vaccines.

We claim: 
     
         1 . A method for identifying a peptide that binds specifically to zona pellucida of oocytes of a target species of animal, the method comprising
 (a) isolating oocytes of the target species of animal and isolating oocytes of one or more non-target species of animals;   (b) contacting the isolated oocytes with a phage library;   (c) selecting phage from the library that bind specifically to the oocytes of the target species of animal, thereby identifying peptides that bind to the zona pellucida of oocytes of the target species of animal.   
     
     
         2 . The method of  claim 1 , wherein the phage library is first contacted with the oocytes of the one of more non-target species of animals, and subsequently, members of the phage library that do not bind to the oocytes of the one of more non-target species of animals are contacted with the oocytes of the target species of animal and those members of the phage library that bind to the oocytes of the target species of animal are selected, thereby identifying peptides that bind specifically to the zona pellucida of oocytes of the target species of animal. 
     
     
         3 . The method of  claim 1 , wherein the phage library is first contacted with the oocytes of the target species of animals, and subsequently, members of the phage library that bind to the oocytes of the target species of animals are contacted with the oocytes of the one or more non-target species of animal and those members of the phage library that do not bind to the oocytes of the one or more non-target species of animal are selected, thereby identifying peptides that bind specifically to the zona pellucida of oocytes of the target species of animal. 
     
     
         4 . The method of  claim 1 , wherein the target species of animal is a swine. 
     
     
         5 . The method of  claim 1 , wherein the non-target species of animals include one or more of felines, canines, and bovines. 
     
     
         6 . An isolated polypeptide comprising an amino acid sequence selected from a group consisting of SEQ ID NO:2 (DANRLPHPANIN), SEQ ID NO: 18(TLGWTANEAPRR), SEQ ID NO: 19 (LLADTTHHRPWT), SEQ ID NO:20(SQSPAMYSQTRP), SEQ ID NO:21 (AVTQHLKFKGFN), and SEQ ID NO:22(ANFNMTHHQGHK). 
     
     
         7 . An isolated polypeptide consisting of an amino acid sequence selected from SEQ ID NO:2 (DANRLPHPANIN), SEQ ID NO: 18 (TLGWTANEAPRR), SEQ ID NO: 19(LLADTTHHRPWT), SEQ ID NO:20 (SQSPAMYSQTRP), SEQ ID NO:21(AVTQHLKFKGFN), and SEQ ID NO:22 (ANFNMTHHQGHK). 
     
     
         8 . An isolated polynucleotide encoding the polypeptide of  claim 6 . 
     
     
         9 . A recombinant polynucleotide comprising a promoter sequence operably linked to the polynucleotide of  claim 8 . 
     
     
         10 . An isolated cell transformed with the recombinant polynucleotide of  claim 9 . 
     
     
         11 . A method of producing a polypeptide encoded by the polynucleotide of  claim 8 , the method comprising:
 a) culturing a cell under conditions suitable for expression of the polypeptide, wherein said cell is transformed with a recombinant polynucleotide, and said recombinant polynucleotide comprises a promoter sequence operably linked to the polynucleotide of  claim 8 ; and   b) recovering the polypeptide so expressed.   
     
     
         12 . A vector comprising the recombinant polynucleotide of  claim 9 . 
     
     
         13 . The vector of  claim 12 , where the vector is a viral vector. 
     
     
         14 . An antigen comprising the polypeptide of  claim 6  conjugated to one or more carrier proteins. 
     
     
         15 . An immunogenic composition comprising;
 (a) a polypeptide comprising an amino acid sequence selected from a group consisting of SEQ ID NO:2 (DANRLPBPANIN), SEQ ID NO: 18 (TLGWTANEAPRR), SEQ ID NO: 19 (LLADTTHHRPWT), SEQ ID NO:20 (SQSPAMYSQTRP), SEQ ID NO:21(AVTQHLKFKGFN), and SEQ ID NO:22 (ANFNMTHHQGHK); and   (b) a suitable excipient, carrier, or diluent.   
     
     
         16 . The composition of  claim 15 , further comprising an adjuvant. 
     
     
         17 . The composition of  claim 16 , comprising two or more polypeptides, each polypeptide comprising an amino acid sequence selected from a group consisting of SEQ ID NO:2 (DANRLPHPANIN), SEQ ID NO:18 (TLGWTANEAPRR), SEQ ID NO:19(LLADTTHHRPWT), SEQ ID NO:20 (SQSPAMYSQTRP), SEQ ID NO:21(AVTQHLKFKGFN), and SEQ ID NO:22 (ANFNMTHHQGHK). 
     
     
         18 . An immunogenic composition comprising:
 (a) the vector of  claim 12 ; and   (b) a suitable excipient, carrier, or diluent.   
     
     
         19 . The composition of  claim 18 , further comprising an adjuvant 
     
     
         20 . A method for producing antibodies that bind to swine sperm, the method comprising administering the immunogenic composition of  claim 15  to an animal. 
     
     
         21 . The method of  claim 20 , wherein the animal is a female swine. 
     
     
         22 . A method for producing antibodies that bind to swine sperm, the method comprising administering the immunogenic composition of  claim 18  to an animal. 
     
     
         23 . The method of  claim 22 , wherein the animal is a female swine. 
     
     
         24 . A method for immunizing a swine against conception, the method comprising administering the immunogenic composition of  claim 15  to the swine. 
     
     
         25 . The method of  claim 24 , wherein the swine is female. 
     
     
         26 . A method for immunizing a swine against conception, the method comprising administering the immunogenic composition of  claim 18  to the swine. 
     
     
         27 . The method of  claim 26 , wherein the swine is female.