Materials and Methods For Cell-Free Expression of Vaccine Epitope Concatemers

Abstract

The present disclosure provides materials and methods for cell-free expression of epitopes for immunotherapy applications. In particular, the present disclosure provides materials and methods for expressing concatenated epitopes using a cell-free protein synthesis platform for high throughput, large scale, and unbiased epitope screening and the generation of multi-epitope vaccines.

What is claimed is: 
     
         1 . An immunogenic vaccine composition comprising two or more concatenated epitopes from at least one pathogenic organism. 
     
     
         2 . The composition of  claim 1 , wherein the two or more concatenated epitopes are selected from a group consisting of: multiple epitopes from a same proteins; multiple epitopes from a different protein; multiple epitopes from the same pathogenic organism; and multiple epitopes from different pathogenic organisms. 
     
     
         3 . The composition of  claim 1 , wherein the two or more concatenated epitopes are capable of generating an immunogenic response in a human subject. 
     
     
         4 . The composition of  claim 1 , wherein the vaccine composition is a modified live vaccine composition, an attenuated vaccine composition, an inactivated vaccine composition, a subunit vaccine composition, or a recombinant vaccine composition. 
     
     
         5 . The composition of  claim 1 , wherein the at least one pathogenic organism is selected from the group consisting of a virus, a parasite, a bacterium, or a fungus. 
     
     
         6 . The composition of  claim 1 , wherein the at least one pathogenic organism is a  Staphylococcus  species. 
     
     
         7 . The composition of  claim 6 , wherein the two or more concatenated epitopes are from fibronectin binding protein-A (FnbA), fibronectin binding protein-B (FnbB), clumping factor A (ClfA), clumping factor B (CHB), serine-rich adhesion for platelets (SraP), collagen adhesin (CNA), elastin binding protein (EbpS), extracellular fibrinogen binding protein (Efb), and enolase (Eno). 
     
     
         8 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO: 1. 
     
     
         9 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise any combination of SEQ ID NOs: 2-8. 
     
     
         10 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO: 9. 
     
     
         11 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise any combination of SEQ ID NOs: 10-16. 
     
     
         12 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO: 17. 
     
     
         13 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise any combination of SEQ ID NOs: 18-24. 
     
     
         14 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO: 25. 
     
     
         15 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise any combination of SEQ ID NOs: 26-33. 
     
     
         16 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise a polypeptide having at least 70% sequence identity to the polypeptide of SEQ ID NO: 34. 
     
     
         17 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise any combination of SEQ ID NOs: 35-41. 
     
     
         18 . The composition of  claim 6 , wherein the two or more concatenated epitopes comprise any combination of SEQ ID NOs: 2-8, 10-16, 18-24, 26-33, and 35-41. 
     
     
         19 . A method of generating an immune response in a human subject, the method comprising:
 administering a therapeutically effective dose of the vaccine composition of  claim 1  to the human subject, wherein the vaccine composition induces an immunogenic response in the human subject against at least one pathogenic organism.   
     
     
         20 . The method of  claim 19 , wherein the administration of the vaccine composition to the human subject reduces the occurrence of at least one symptom caused by the at least one pathogenic organism. 
     
     
         21 . The method of  claim 19 , wherein the at least one pathogenic organism is selected from the group consisting of a virus, a parasite, a bacterium, or a fungus. 
     
     
         22 . The method of  claim 19 , wherein the at least one pathogenic organism is a  Staphylococcus  species. 
     
     
         23 . The method of  claim 20 , wherein the at least one symptom comprises a skin infection, a bloodstream infection, pneumonia, a bone infection, a joint infection, and/or endocarditis. 
     
     
         24 . The method of  claim 19 , wherein the vaccine composition is administered to the human subject prior to exposure to the at least one pathogenic organism.