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 . (canceled) 
     
     
         3 . The composition of  claim 1 , wherein said bacteriophage is bacteriophage Qβ. 
     
     
         4 - 6 . (canceled) 
     
     
         7 . The composition of  claim 1 , wherein said virus-like particle of an RNA bacteriophage comprises recombinant coat proteins having an amino acid sequence of SEQ ID NO:159, or a mixture of coat proteins having amino acid sequences of SEQ ID NO:159 and of SEQ ID NO:217. 
     
     
         8 - 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, and wherein said amino acid linker comprises said second attachment site. 
     
     
         14 - 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 asthma,   (e) a recombinant protein of breast cancer cells,   (f) a recombinant protein of kidney cancer cells,   (g) a recombinant protein of prostate cancer cells,   (h) a recombinant protein of skin cancer cells,   (i) a recombinant protein of brain cancer cells,   (j) a recombinant protein of leukemia cells,   (k) a recombinant protein of Influenza virus,   (l) a recombinant protein of HIV,   (m) a recombinant protein of Hepatitis C virus,   (n) a recombinant protein of  Toxoplasma,      (O) a recombinant protein of  Plasmodium falciparum,      (p) a recombinant protein of  Plasmodium vivax,      (q) a recombinant protein of  Plasmodium ovale,      (r) a recombinant protein of  Plasmodium malariae , or   (s) 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. 
     
     
         35 . The composition of  claim 34 , wherein said infectious disease of viral etiology is selected from the group consisting of:
 (a) HIV,   (b) influenza,   (c) herpes,   (d) viral hepatitis,   (e) Epstein Bar, and   (f) viral encephalitis.   
     
     
         36 . (canceled) 
     
     
         37 . (canceled) 
     
     
         38 . The composition of  claim 1 , wherein said antigen is suitable to prevent or treat an infectious disease of parasitic etiology. 
     
     
         39 . The composition of  claim 38 , wherein said infectious disease of parasitic etiology is malaria, trypanosomiasis, leishmaniasis, trichommoniasis or amoebiasis. 
     
     
         40 . The composition of  claim 1 , wherein said antigen or antigenic determinant is a peptide, a protein, or a fragment of a protein, selected from the group consisting of:
 (a) an Influenza M2 protein;   (b) a phospholipase A2 protein; and   (c) a Der p I peptide.   
     
     
         41 - 43 . (canceled) 
     
     
         44 . The composition of  claim 1 , wherein said antigen or antigenic determinant is a peptide consisting of the amino acids 2 to 24 of SEQ ID NO: 213 or a fragment thereof being at least 6 amino acids in length. 
     
     
         45 - 47 . (canceled) 
     
     
         48 . The composition of  claim 1 , wherein said antigen or antigenic determinant is a phospholipase A2 protein or fragment thereof comprising an amino acid sequence selected from the group consisting of:
 (a) the amino acid sequence of SEQ ID NO:168;   (b) the amino acid sequence of SEQ ID NO: 169;   (c) the amino acid sequence of SEQ ID NO:170;   (d) the amino acid sequence of SEQ ID NO: 171;   (e) the amino acid sequence of SEQ ID NO:172;   (f) the amino acid sequence of SEQ ID NO:173;   (g) the amino acid sequence of SEQ ID NO:174; and   (h) the amino acid sequence of SEQ ID NO:175.   
     
     
         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 . (canceled) 
     
     
         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 recombinant protein or antigenic fragment thereof, of Influenza virus. 
     
     
         55 . The composition of  claim 1 , wherein said antigen or antigenic determinant is influenza antigen hemagglutinin or an antigenic fragment thereof. 
     
     
         56 - 65 . (canceled) 
     
     
         66 . The composition of  claim 1 , wherein said antigen or antigenic determinant is a self antigen, or antigenic fragment thereof, 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. 
     
     
         67 . (canceled) 
     
     
         68 . The composition of  claim 66 , wherein said self antigen is a human IgE. 
     
     
         69 . The composition of  claim 66 , wherein said self antigen is Interleukin-17. 
     
     
         70 . The composition of  claim 69 , wherein said hetero bifunctional linker is succinimidyl-6-(β-maleimidopropionamid)hexanoate (SMPH). 
     
     
         71 . The composition of  claim 70 , wherein said Interleukin-17 comprises amino acids 24-155 of SEQ ID NO: 228. 
     
     
         72 . The composition of  claim 71 , wherein said composition further comprises an amino acid linker that is bound to said antigen or said antigenic determinant by way of at least one covalent peptide bond, and wherein said amino acid linker comprises said second attachment site. 
     
     
         73 . The composition of  claim 72 , wherein said amino acid linker is GGC. 
     
     
         74 . The composition of  claim 73 , wherein said amino acid linker is fused to the C-terminus of said Interleukin-17. 
     
     
         75 . (canceled) 
     
     
         76 . The composition of  claim 68 , wherein said heterobifunctional linker is SMPH. 
     
     
         77 . The composition of  claim 1 , wherein said virus-like particle of RNA bacteriophage is a recombinant virus-like particle of RNA bacteriophage Qβ coat protein, and wherein said RNA bacteriophage Qβ coat protein consists of the amino acid sequence of SEQ ID NO:159. 
     
     
         78 . The composition of  claim 31 , wherein said virus-like particle of RNA bacteriophage is a recombinant virus-like particle of RNA bacteriophage Qβ coat protein, and wherein said RNA bacteriophage Qβ coat protein consists of the amino acid sequence of SEQ ID NO:159. 
     
     
         79 - 83 . (canceled) 
     
     
         84 . The composition of  claim 76 , wherein said virus-like particle of RNA bacteriophage is a recombinant virus-like particle of RNA bacteriophage Qβ coat protein, and wherein said RNA bacteriophage Qβ coat protein consists of the amino acid sequence of SEQ ID NO:159.