Rsv f protein compositions and methods for making same

Abstract

The present invention relates to immunogenic compositions comprising RSV F protein, methods for preparing compositions that contain RSV F protein ecto-domain polypeptides, and to certain engineered RSV F proteins and nucleic acids that encode the engineered RSV F proteins. Compositions prepared using the methods can contain RSV F protein ecto-domain polypeptides in a predominant or single desired form and conformation. The invention also relates to methods for inducing an immune response to RSV F.

1 .- 6 . (canceled) 
     
     
         7 . A respiratory syncytial virus (RSV)-F protein comprising a F1 fragment, a F2 fragment, and an oligomerization domain,
 wherein the F1 fragment and the F2 fragment comprise an amino acid sequence corresponding to the amino acid sequence of the F1 fragment and the F2 fragment set forth in SEQ ID NO: 1 or SEQ ID NO: 2;   wherein the F2 fragment lacks a transmembrane domain and a cytoplasmic tail;   wherein the oligomerization domain is a trimerizing sequence from bacteriophage T4 fibritin; and   wherein 1 to 30 of the amino acid residues in the RSV-F protein are substituted with another amino acid residue as compared to the amino acid sequence set forth in SEQ ID NO: 1 or SEQ ID NO: 2.   
     
     
         8 . The RSV-F protein of  claim 7 , wherein the RSV-F protein does not comprise a p27 region. 
     
     
         9 . The RSV-F protein of  claim 7 , wherein the trimerizing sequence comprises the amino acid sequence of SEQ ID NO: 19. 
     
     
         10 . The RSV-F protein of  claim 7 , wherein the trimerizing sequence comprises amino acid residues 3-29 of SEQ ID NO: 19. 
     
     
         11 . The RSV-F protein of  claim 7 , wherein 1 to 15 of the amino acid residues in the RSV-F protein are substituted with another amino acid residue as compared to the amino acid sequence set forth in SEQ ID NO: 1 or SEQ ID NO: 2. 
     
     
         12 . A nucleic acid encoding the RSV-F protein of  claim 7 . 
     
     
         13 . A vector comprising the nucleic acid of  claim 12 . 
     
     
         14 . A host cell comprising the vector of  claim 13 . 
     
     
         15 . A method of making a respiratory syncytial virus (RSV)-F protein comprising a F1 fragment, a F2 fragment, and an oligomerization domain, wherein the F1 fragment and the F2 fragment comprise an amino acid sequence corresponding to the amino acid sequence of the F1 fragment and the F2 fragment set forth in SEQ ID NO: 1 or SEQ ID NO: 2; wherein the F2 fragment lacks a transmembrane domain and a cytoplasmic tail; wherein the oligomerization domain is a trimerizing sequence from bacteriophage T4 fibritin; and wherein 1 to 30 of the amino acid residues in the RSV-F protein are substituted with another amino acid residue as compared to the amino acid sequence set forth in SEQ ID NO: 1 or SEQ ID NO: 2, the method comprising culturing the host cell of  claim 14  and recovering the RSV-F protein from a culture media. 
     
     
         16 . An immunogenic composition comprising the RSV-F protein of  claim 7 . 
     
     
         17 . The immunogenic composition of  claim 16 , further comprising an adjuvant. 
     
     
         18 . A method of inducing an immune response to RSV in a subject in need thereof, the method comprising administering a therapeutically effective amount of the immunogenic composition of  claim 16  to the subject.