Methods to Determine the Presence of an Antibody that Binds Modified Human Thymic Stromal Lymphopoietin (TSLP)

Abstract

Modified, furin resistant human TSLP polypeptides and polynucleotides encoding the modified human TSLP polypeptides are provided. Pharmaceutical compositions, B and T cell activation agents, assays and methods of use are also described.

We claim: 
     
         1 . An isolated nucleic acid encoding a polypeptide comprising at least 90% amino acid sequence identity to amino acids 29-159 of SEQ ID NO: 10, wherein the polypeptide comprises one or more amino acid substitutions or deletions to inactivate the furin cleavage site RRKRK (SEQ ID NO:6) at position 127-131 of SEQ ID NO:4. 
     
     
         2 . The nucleic acid of  claim 1  encoding a polypeptide comprising at least 90% amino acid sequence identity to SEQ ID NO: 12, wherein the polypeptide comprises one or more amino acid substitutions or deletions to inactivate the furin cleavage site RRKRK (SEQ ID NO:6) at position 127-131 of SEQ ID NO:4. 
     
     
         3 . The nucleic acid of  claim 1  encoding a polypeptide comprising at least 90% amino acid sequence identity to SEQ ID NO: 14, wherein the polypeptide comprises one or more amino acid substitutions or deletions to inactivate the furin cleavage site RRKRK (SEQ ID NO:6) at position 127-131 of SEQ ID NO:4. 
     
     
         4 . The nucleic acid of  claim 1  encoding a polypeptide comprising at least 90% amino acid sequence identity to SEQ ID NO: 16, wherein the polypeptide comprises one or more amino acid substitutions or deletions to inactivate the furin cleavage site RRKRK (SEQ ID NO:6) at position 127-131 of SEQ ID NO:4. 
     
     
         5 . The nucleic acid of  claim 1  encoding a polypeptide comprising at least 90% amino acid sequence identity to SEQ ID NO: 17, wherein the polypeptide comprises an amino acid substitution or deletion to inactivate the furin cleavage site RRKRK (SEQ ID NO:6) at position 127-131 of SEQ ID NO:4. 
     
     
         6 . The nucleic acid of  claim 1  encoding a polypeptide comprising at least 90% amino acid sequence identity to SEQ ID NO: 18, wherein the polypeptide comprises one or more amino acid substitutions or deletions to inactivate the furin cleavage site RRKRK (SEQ ID NO:6) at position 127-131 of SEQ ID NO:4. 
     
     
         7 . The nucleic acid molecule of  claim 1 , wherein the nucleic acid comprises a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 9, 11, 13 and 15. 
     
     
         8 . The nucleic acid molecule of  claim 1 , further comprising a nucleotide sequence encoding a heterologous protein in frame with the polynucleotide of  claim 1 . 
     
     
         9 . The nucleic acid molecule of  claim 8 , wherein the heterologous protein is a cell targeting moiety. 
     
     
         10 . The nucleic acid molecule of  claim 9 , wherein the cell targeting moiety is an antibody that binds a cell surface antigen. 
     
     
         11 . The nucleic acid molecule of  claim 9 , wherein the cell targeting moiety is a ligand that binds a cell surface receptor. 
     
     
         12 . The nucleic acid molecule of  claim 8 , wherein the heterologous protein is a peptide tag. 
     
     
         13 . The nucleic acid molecule of  claim 8 , wherein the heterologous protein is an Fc polypeptide. 
     
     
         14 . The nucleic acid molecule of  claim 1 , operably linked to a transcriptional or translational regulatory sequence. 
     
     
         15 . The nucleic acid molecule of  claim 14 , wherein said transcriptional or translational regulatory sequence comprise a transcriptional promoter or enhancer. 
     
     
         16 . An isolated vector comprising the nucleic acid molecule of  claim 1 . 
     
     
         17 . An isolated host cell comprising the nucleic acid molecule of  claim 1 . 
     
     
         18 . The isolated host cell of  claim 17 , wherein the host cell is a mammalian cell.