Polypeptide comprising the amino acid of an N-terminal choline binding protein a truncate, vaccine derived therefrom and uses thereof

Abstract

This invention provides an isolated polypeptide comprising an amino acid sequence of a N-terminal choline binding protein A truncate in which the amino acid sequence is set forth in any of SEQ ID NOS: 1, 3-7, or 9-11, including fragments, mutants, variants, analogs, or derivatives, thereof. Also, this invention provides a isolated polypeptide comprising an amino acid sequence of a N-terminal choline binding protein A truncate, wherein the amino acid is set forth in SEQ ID NO: 24, wherein the polypeptide retains its native tertiary structure and methods of preparation. This invention provides an isolated polypeptide comprising an amino acid sequence of a N-terminal choline binding protein A truncate, wherein the polypeptide has lectin activity and does not bind to choline. This invention provides an isolated immunogenic polypeptide comprising an amino acid sequence of a N— terminal choline binding protein A truncate. This invention provides an isolated nucleic acid encoding a polypeptide comprising an amino acid sequence of a N— terminal choline binding protein A truncate. Lastly, this invention provides pharmaceutical compositions, vaccines, and diagnostic and therapeutic methods of use.

1 - 45 . (canceled)  
     
     
         46 . An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of: 
 a) a nucleotide sequence encoding a polypeptide comprising a variant of SEQ ID NO: 1, 3, or 24, wherein said variant comprises at least one to 57 amino acid substitutions and said polypeptide does not bind to choline and is immunogenic;    b) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence set forth in set forth in SEQ ID NO: 1, 3, 4, 5, 22, or 24, wherein said polypeptide does not bind to choline;    c) a nucleotide sequence encoding a polypeptide comprising a variant of SEQ ID NO:4, 5, or 22, wherein said variant comprises at least one to 57 amino acid substitutions, does not bind to choline, is immunogenic, and comprises up to 398 amino acids;    d) a nucleotide sequence encoding a polypeptide comprising at least 138 consecutive amino acids of SEQ ID NO:24, wherein said polypeptide does not bind to choline;    e) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence set forth in SEQ ID NO:7 or 9, wherein said polypeptide comprises up to 376 amino acids and does not bind to choline;    f) a nucleotide sequence encoding a polypeptide comprising a variant of SEQ ID NO:7 or 9, wherein said variant comprises at least one to 57 amino acid substitutions, comprises up to 376 amino acids, does not bind choline, and is immunogenic;    g) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence set forth in SEQ ID NO:10, 11, or 23, wherein said polypeptide comprises up to 328 amino acid and does not bind choline;    h) a nucleotide sequence encoding a polypeptide comprising a variant of SEQ ID NO:10, 11, or 23, wherein said variant comprises at least one to 57 amino acid substitutions, comprises up to 328 amino acids, does not bind choline, and is immunogenic; and,    i) a nucleotide sequence comprises SEQ ID NO: 15, 12, 14, 16, 17, 19, 20, or 21 wherein, said nucleotide sequence encodes a polypeptide that does not bind choline.    
     
     
         47 . The isolated nucleic acid molecule of  claim 46 , wherein said nucleic acid molecule is operably linked to a promoter of RNA transcription.  
     
     
         48 . The isolated nucleic acid molecule of  claim 46 , wherein the nucleic acid molecule is DNA or RNA.  
     
     
         49 . The isolated nucleic acid molecule of  claim 46 , wherein the nucleic acid molecule is cDNA or genomic DNA.  
     
     
         50 . A vector comprising the nucleic acid molecule of  claim 46 .  
     
     
         51 . The vector of  claim 50 , wherein said nucleic acid sequence is operably linked to a promoter that regulates transcription.  
     
     
         52 . The vector of  claim 50 , wherein the vector is a plasmid, a cosmid, a yeast artificial chromosome (YAC), a bacteriophage, or eukaryotic viral DNA.  
     
     
         53 . A method of obtaining a polypeptide in purified form comprising: 
 a) introducing the vector of  claim 50  into a suitable host cell;    b) culturing the resulting host cell so as to produce the polypeptide;    c) recovering the polypeptide produced in step (b); and,    d) purifying the polypeptide so recovered in step (c).    
     
     
         54 . A vaccine comprising an isolated nucleic acid molecule encoding a polypeptide and a pharmaceutically acceptable adjuvant or carrier, wherein said polypeptide comprises an amino acid sequence of an N-terminal choline binding protein A truncate that does not bind to choline.  
     
     
         55 . The vaccine of  claim 54 , wherein said nucleic acid molecule comprises the nucleotide sequence of  claim 46 .  
     
     
         56 . A pharmaceutical composition comprising an isolated antibody that binds to a polypeptide comprising an amino acid sequence of an N-terminal choline binding protein A truncate, wherein said isolated antibody does not bind to a choline binding domain and said isolated antibody is substantially free of antibodies that bind to the choline binding domain.  
     
     
         57 . The pharmaceutical composition of  claim 56 , wherein said amino acid sequence comprises SEQ ID NO:7, 9, 10, 11, 23, 1, 3, 4, 5, 22, or 24.  
     
     
         58 . The pharmaceutical composition of  claim 56 , wherein the antibody is a monoclonal antibody.  
     
     
         59 . The pharmaceutical composition of  claim 56 , wherein the antibody is a polyclonal antibody.  
     
     
         60 . A method for inducing an immune response in a subject which has been exposed to or infected with a pneumococcal bacterium comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising the vaccine of  claim 54 , thereby inducing an immune response.  
     
     
         61 . A method for preventing infection by a pneumococcal bacterium in a subject comprising administering to the subject an amount of a pharmaceutical composition comprising a polypeptide having an amino acid sequence of an N-terminal choline binding protein A truncate which does not bind choline, wherein said amount is effective to prevent pneumococcal bacterium attachment, thereby preventing infection by a pneumococcal bacterium.  
     
     
         62 . The method of  claim 61 , wherein said polypeptide comprises an amino acid sequence selected from the group consisting of: 
 a) an amino acid sequence set forth in SEQ ID NO:1, 4, 3, 5, 22, 24, 7, 9, 10, 11, or 23, wherein polypeptide does not bind choline; and,    b) a variant of an amino acid sequence set forth in SEQ ID NO: 1, 4, 3, 5, 22, 24, 7, 9, 10, 11, or 23, wherein said variant comprises at least one to 57 amino acid substitutions.    
     
     
         63 . A method for preventing infection by a pneumococcal bacterium in a subject comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition of  claim 56 , and thereby preventing an infection by the pneumococcal bacterium.  
     
     
         64 . A method for treating a subject infected with or exposed to pneumococcal bacterium comprising administering to the subject a therapeutically effective amount of a vaccine of  claim 54 , thereby treating the subject.