Molecular antigen arrary

Abstract

The present invention is related to the fields of molecular biology, virology, immunology and medicine. The invention provides a composition comprising an ordered and repetitive antigen or antigenic determinant array. The invention also provides a process for producing an antigen or antigenic determinant in an ordered and repetitive array. The ordered and repetitive antigen or antigenic determinant is useful in the production of vaccines for the treatment of infectious diseases, the treatment of allergies and as a pharmaccine to prevent or cure cancer and to efficiently induce self-specific immune responses, in particular antibody responses.

1 - 50 . (canceled) 
     
     
         51 . A method of immunizing an animal comprising administering to an animal a composition, which composition comprises:
 (a) a non-natural molecular scaffold comprising:
 (i) a core particle that is a virus-like particle of an RNA bacteriophage; and 
 (ii) an organizer comprising at least one first attachment site, wherein said organizer is connected to said core particle by at least one covalent bond; and 
   (b) an antigen or antigenic determinant with at least one second attachment site,
 wherein said antigen or antigenic determinant is amyloid beta peptide (Aβ 1-42  or a fragment thereof, and wherein said second attachment site is selected from the group consisting of: 
 (i) an attachment site not naturally occurring with said antigen or antigenic determinant; and 
 (ii) an attachment site naturally occurring with said antigen or antigenic determinant, 
   wherein said second attachment site associates with said first attachment site through at least one non-peptide bond; and   wherein said antigen or antigenic determinant and said non-natural molecular scaffold interact through said association to form an ordered and repetitive antigen array,   wherein an immune response against said antigen or antigenic determinant is produced in said animal.   
     
     
         52 - 53 . (canceled) 
     
     
         54 . The method of  claim 51 , wherein said association is by way of at least one covalent non-peptide bond. 
     
     
         55 . The method of  claim 51 , wherein said virus-like particle of an RNA bacteriophage comprises recombinant proteins, or fragments thereof, of an RNA bacteriophage. 
     
     
         56 . The method of  claim 51 , wherein said RNA bacteriophage is selected from the group consisting of:
 (a) bacteriophage Qβ;   (b) bacteriophage R17;   (c) bacteriophage fr;   (d) bacteriophage GA;   (e) bacteriophage SP;   (f) bacteriophage MS2;   (g) bacteriophage M11;   (h) bacteriophage MX1;   (i) bacteriophage NL95;   (k) bacteriophage f2; and   (l) bacteriophage PP7.   
     
     
         57 . The method of  claim 51 , wherein said virus-like particle of an RNA bacteriophage comprises recombinant proteins, or fragments thereof, of bacteriophage Qβ. 
     
     
         58 . The method of  claim 57 , wherein said second attachment site does not naturally occur with said antigen or antigenic determinant, and wherein said composition comprises an amino acid linker bound to said antigen or antigenic determinant through at least one covalent peptide bond, and wherein said amino acid linker comprises said second attachment site. 
     
     
         59 . The method of  claim 58 , wherein said amino acid linker comprises a sulfhydryl group. 
     
     
         60 . The method of  claim 58 , wherein said amino acid linker comprises a cysteine residue. 
     
     
         61 . The method of  claim 58 , wherein said amino acid linker is GGC or GGC-NH 2 . 
     
     
         62 . The composition of  claim 51 , wherein said amyloid beta peptide (Aβ 1-42 ) or a fragment thereof is selected from the group consisting of:
 (a) Aβ 1-15;   (b) Aβ 1-27;   (c) Aβ 1-40;   (d) Aβ 1-42;   (e) Aβ 33-40; and   (e) Aβ 33-42.   
     
     
         63 . The method of  claim 51 , wherein said amyloid beta peptide (Aβ 1-42 ) or a fragment thereof with said second attachment site has an amino acid sequence selected from the group consisting of: 
       
         
           
                 
                 
               
                   (SEQ ID NO:367) 
                     
                 
                 
                 
                 
               
                     
                   (a) DAEFRHDSGYEVHHQGGC; 
                     
                 
                     
                     
                 
                 
                 
               
                   (SEQ ID NO:369) 
                     
                 
                 
                 
                 
               
                     
                   (b) CGHGNKSGLMVGGVVIA; 
                     
                 
                     
                   and 
                 
                     
                     
                 
                 
                 
               
                   (SEQ ID NO:368) 
                     
                 
                 
                 
                 
               
                     
                   (c) DAEFRHDSGYEVHHQKLVF FAEDVGSNGGC. 
                     
                 
             
                
               
            
             
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
               
            
           
         
       
     
     
         64 . The method of  claim 51 , wherein said organizer is a polypeptide or residue thereof and said second attachment site is a polypeptide or residue thereof. 
     
     
         65 . The method of  claim 51 , wherein said composition further comprises a heterobifunctional cross-linker. 
     
     
         66 . The method of  claim 65 , wherein said heterobifunctional cross-linker is selected from the group consisting of:
 (a) SMPH;   (b) Sulfo-MBS; and   (c) Sulfo-GMBS.   
     
     
         67 . The method of  claim 51 , wherein said first attachment site comprises an amino group and said second attachment site comprises a sulfhydryl group. 
     
     
         68 . The method of  claim 51 , wherein said first attachment site comprises a lysine residue and said second attachment site comprises a cysteine residue. 
     
     
         69 . The method of  claim 51 , wherein said first attachment site comprises an amino group or a lysine residue. 
     
     
         70 . The method of  claim 51 , wherein said second attachment site comprises a sulfhydryl group or a cysteine residue. 
     
     
         71 . The method of  claim 51 , wherein said first attachment site is not a sulfhydryl group. 
     
     
         72 . The method of  claim 68 , wherein said RNA bacteriophage is bacteriophage Qβ. 
     
     
         73 . The method of  claim 51 , wherein said second attachment site does not naturally occur with said antigen or antigenic determinant. 
     
     
         74 . The method of  claim 51 , wherein said virus-like particle of an RNA bacteriophage comprises recombinant coat proteins comprising an amino acid sequence selected from the group consisting of:
 (a) SEQ ID NO:159;   (b) SEQ ID NO:160;   (c) SEQ ID NO:161;   (d) SEQ ID NO:162;   (e) SEQ ID NO:163;   (f) SEQ ID NO:164;   (g) SEQ ID NO:165;   (h) SEQ ID NO:166;   (i) SEQ ID NO:167;   (j) SEQ ID NO:215;   (k) SEQ ID NO:253;   (l) SEQ ID NO:217; and   (m) SEQ ID NO:254.   
     
     
         75 . The method of  claim 51 , wherein said virus-like particle of an RNA bacteriophage comprises recombinant coat proteins having the amino acid sequence of SEQ ID NO:159. 
     
     
         76 . The method of  claim 51 , wherein said virus-like particle of an RNA bacteriophage consists essentially of coat proteins having the amino acid sequence of SEQ ID NO:159. 
     
     
         77 . The method of  claim 51 , wherein said virus-like particle of an RNA bacteriophage comprises one or more coat proteins of said RNA bacteriophage that have been modified by deletion or substitution to remove at least one naturally occurring lysine residue, or that have been modified by insertion or substitution to add at least one lysine residue. 
     
     
         78 . The method of  claim 77 , wherein said RNA bacteriophage is Qβ. 
     
     
         79 . The method of  claim 51 , wherein said virus-like particle of an RNA bacteriophage comprises one or more coat proteins comprising an amino acid sequence selected from the group consisting of:
 (a) SEQ ID NO:255;   (b) SEQ ID NO:256;   (c) SEQ ID NO:257;   (d) SEQ ID NO:258;   (e) SEQ ID NO:259; and   (f) a mixture of any one of (a)-(e) and the corresponding A1 protein.   
     
     
         80 . The method of  claim 51 , wherein said organizer is an integral part of said RNA bacteriophage. 
     
     
         81 . The method of  claim 51 , wherein said virus-like particle is a recombinant virus-like particle. 
     
     
         82 . The method of  claim 72 , wherein said association is by way of at least one covalent non-peptide bond. 
     
     
         83 . The method of  claim 82 , wherein said virus-like particle of an RNA bacteriophage comprises recombinant coat proteins having the amino acid sequence of SEQ ID NO:159. 
     
     
         84 . The method of  claim 83 , wherein said composition further comprises a heterobifunctional cross-linker selected from the group consisting of:
 (a) SMPH;   (b) Sulfo-MBS; and   (c) Sulfo-GMBS.