Molecular Antigen Array

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 . A composition comprising:
 (a) a non-natural molecular scaffold comprising:
 (i) a core particle which is a virus-like particle of an RNA bacteriophage comprising at least one recombinant protein of said RNA bacteriophage; and 
 (ii) an organizer comprising at least one first attachment site,
 wherein said organizer is an integral part of said core particle and is connected to said core particle by at least one covalent bond; and wherein said first attachment site is an amino group of a lysine residue of said recombinant protein of said RNA bacteriophage; and 
 
   (b) at least one antigen or antigenic determinant with at least one second attachment site, wherein said antigen or antigenic determinant is not amyloid β or a peptide or fragment thereof, and wherein said second attachment site associates with said first attachment site through a heterobifunctional linker via at least one non-peptide covalent bond; and wherein said second attachment site is a sulfhydryl group of a cysteine residue, and wherein said antigen or antigenic determinant and said scaffold interact through said association to form an ordered and repetitive antigen array.   
     
     
         2 . The composition of  claim 1 , 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;   (j) bacteriophage f2; and   (k) bacteriophage PP7.   
     
     
         3 . The composition of  claim 1 , wherein said bacteriophage is bacteriophage Qβ. 
     
     
         4 . (canceled) 
     
     
         5 . (canceled) 
     
     
         6 . The composition of  claim 1 , 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.   
     
     
         7 .- 12 . (canceled) 
     
     
         13 . The composition of  claim 1  further comprising an amino acid linker, wherein said amino acid linker is bound to said antigen or said antigenic determinant by way of at least one covalent bond, wherein said covalent bond is a peptide bond. 
     
     
         14 . (canceled) 
     
     
         15 . (canceled) 
     
     
         16 . The composition of  claim 13 , wherein said amino acid linker comprises said second attachment site. 
     
     
         17 .- 21 . (canceled) 
     
     
         22 . The composition of  claim 1 , wherein said heterobifunctional linker is selected from the group consisting of:
 (a) a maleimidocaproic acid N-hydroxysuccinimide ester;   (b) N-Succinimidyl 3-(2-pyridyldithio) propionate (SPDP); and   (c) Sulfo-MBS.   
     
     
         23 .- 26 . (canceled) 
     
     
         27 . The composition of  claim 1 , wherein said second attachment site does not naturally occur within said antigen or antigenic determinant. 
     
     
         28 .- 30 . (canceled) 
     
     
         31 . The composition of  claim 1 , wherein only one of said second attachment sites associates with said first attachment site through at least one non-peptide covalent bond leading to a single and uniform type of binding of said antigen to said core particle, wherein said only one second attachment site that associates with said first attachment site is a sulfhydryl group. 
     
     
         32 . The composition of  claim 1 , wherein said antigen is a protein or a fragment thereof, being selected from the group consisting of:
 (a) proteins suitable to induce an immune response against allergens,   (b) proteins suitable to induce an immune response against cancer cells,   (c) proteins suitable to induce an immune response against infectious diseases, and   (d) proteins suitable to induce an immune response in farm animals or pets.   
     
     
         33 . The composition of  claim 1 , wherein said antigen is:
 (a) a recombinant protein of bee sting allergy,   (b) a recombinant protein of nut allergy,   (c) a recombinant protein of food allergies,   (d) a recombinant protein of breast cancer cells,   (e) a recombinant protein of kidney cancer cells,   (f) a recombinant protein of prostate cancer cells,   (g) recombinant protein of skin cancer cells,   (h) a recombinant protein of brain cancer cells,   (i) a recombinant protein of leukemia cells,   (j) a recombinant protein of Influenza virus,   (k) a recombinant protein of HIV,   (l) a recombinant protein of Hepatitis virus,   (m) a recombinant protein of  Toxoplasma,      (n) a recombinant protein of  Plasmodium falciparum,      (o) a recombinant protein of  Plasmodium vivax,      (p) a recombinant protein of  Plasmodium ovale,      (q) a recombinant protein of  Plasmodium malariae , or   (r) a recombinant protein of  Chlamydia.      
     
     
         34 . The composition of  claim 1 , wherein said antigen is suitable to prevent or treat an infectious disease of viral etiology, wherein said infectious disease of viral etiology is selected from the group consisting of:
 (a) HIV,   (b) influenza,   (c) viral hepatitis,   (e) Epstein Bar, and   (f) viral encephalitis.   
     
     
         35 . (canceled) 
     
     
         36 . The composition of  claim 1 , wherein said antigen is suitable to prevent or treat an infectious disease of bacterial etiology, wherein said infectious disease of bacterial etiology is a pneumonia, syphillis or tuberculosis. 
     
     
         37 . (canceled) 
     
     
         38 . The composition of  claim 1 , wherein said antigen is suitable to prevent or treat an infectious disease of parasitic etiology, wherein said infectious disease of parasitic etiology is malaria, trypanosomiasis, leishmaniasis, trichomoniasis or amoebiasis. 
     
     
         39 .- 48 . (canceled) 
     
     
         49 . A pharmaceutical composition comprising:
 (a) the composition of  claim 1 ; and   (b) an acceptable pharmaceutical carrier.   
     
     
         50 . A method of immunization of an animal comprising administering to said animal the composition of  claim 1 , wherein an immune response against said antigen or antigenic determinant is produced in said animal. 
     
     
         51 . An immunogenic composition comprising the composition of  claim 1  and an adjuvant. 
     
     
         52 . A process for producing a non-naturally occurring, ordered and repetitive antigen array comprising:
 (a) providing a non-natural molecular scaffold comprising:
 (i) a core particle comprising a virus-like particle of an RNA bacteriophage; and 
 (ii) an organizer which is at least one first attachment site, 
   
       wherein said organizer is an integral part of said core particle and is connected to said core particle by at least one covalent bond; and wherein said first attachment site is an amino group; and
 (b) providing at least one antigen or antigenic determinant with at least one second attachment site, wherein said antigen or antigenic determinant is not amyloid β or a peptide or fragment thereof, and wherein said second attachment site site associates with said first attachment site through a heterobifunctional linker via at least one non-peptide covalent bond; and wherein said second attachment site is a sulfhydryl group, and 
 (c) combining said non-natural molecular scaffold and said antigen to form an ordered and repetitive antigen array. 
 
     
     
         53 . The composition of  claim 1 , wherein said virus-like particle of an RNA bacteriophage comprises one or more recombinant coat proteins consisting of the amino acid sequence of SEQ ID NO:159. 
     
     
         54 . The composition of  claim 1 , wherein said antigen or antigenic determinant is a protein, or an antigenic fragment thereof, suitable to induce an immune response against cancer. 
     
     
         55 . The composition of  claim 1 , wherein said antigen or antigenic determinant is a protein, a peptide or a fragment thereof, selected from
 (a) Her2,   (b) GD2,   (c) EGF-R,   (d) CEA,   (e) CD52,   (f) human melanoma protein gp100,   (g) human melanoma protein melan-A/MART-1,   (h) tyrosinase,   (i) NA17-A nt protein,   (j) MAGE-3 protein,   (k) p53 protein, and   (l) HPV16 E7 protein.   
     
     
         56 . The composition of  claim 1 , wherein said antigen or antigenic determinant is a protein, a peptide or a fragment thereof, selected from
 (a) CCL21,   (b) CXCL12, and   (c) M-CSF.