Polynucleotides encoding E38N interferon gamma polypeptides

Abstract

A conjugate exhibiting interferon gamma activity and comprising at least one first non-polypeptide moiety covalently linked to an IFG polypeptide, the polypeptide comprising an amino acid sequence that differs from that of a parent IFNG polypeptide in at least one introduced and/or at least one removed amino acid residue comprising an attachment group for the non-polypeptide moiety. The conjugate may be used for treatment of various diseases.

1 - 35 . (canceled) 
     
     
         36 . A polynucleotide encoding an E38N interferon gamma (IFNG) polypeptide variant exhibiting IFNG receptor binding activity, said encoded E38N IFNG polypeptide variant having an amino acid sequence which
 (i) varies by 1 to 15 amino acid residue differences from the wild-type human IFNG polypeptide sequence shown in SEQ ID NO: 2 or from a C-terminally truncated fragment thereof wherein said fragment is C-terminally truncated by deletion of the last 1 to 15 amino acid residues relative to SEQ ID NO: 2; and   (ii) is characterized by the substitution E38N.   
     
     
         37 . The polynucleotide of  claim 36 , wherein the 1 to 15 amino acid residue differences in the encoded E38N IFNG polypeptide variant comprises the substitution S40T. 
     
     
         38 . The polynucleotide of  claim 36 , wherein the encoded E38N IFNG polypeptide variant has an amino acid sequence that varies by 1 to 8 amino acid residue differences from the wild-type human IFNG sequence shown in SEQ ID NO: 2 or from said C-terminally truncated fragment thereof. 
     
     
         39 . The polynucleotide of  claim 38 , wherein the encoded E38N IFNG polypeptide variant sequence varies by 1 to 5 amino acid residue differences from the wild-type human IFNG sequence shown in SEQ ID NO: 2 or from said C-terminally truncated fragment thereof. 
     
     
         40 . The polynucleotide of  claim 36 , wherein the encoded E38N IFNG polypeptide variant is C-terminally truncated by 11 amino acid residues. 
     
     
         41 . The polynucleotide of  claim 36 , wherein the encoded E38N IFNG polypeptide variant has a glycosylation site. 
     
     
         42 . The polynucleotide of  claim 41 , wherein the encoded E38N IFNG polypeptide variant has glycosylation sites at residues N25, N38 and N97. 
     
     
         43 . The polynucleotide of  claim 36 , wherein the encoded E38N IFNG polypeptide variant differs by 2-5 amino acid substitutions relative to the wild-type human IFNG sequence shown in SEQ ID NO: 2 or said C-terminally truncated fragment thereof. 
     
     
         44 . The polynucleotide of  claim 43 , wherein the encoded E38N IFNG polypeptide variant wherein said 2-5 substitutions comprise the substitution S40T. 
     
     
         45 . The polynucleotide of  claim 44 , wherein the encoded E38N IFNG polypeptide variant is also C-terminally truncated by 11 amino acid residues. 
     
     
         46 . The polynucleotide of  claim 45 , wherein the encoded E38N IFNG polypeptide variant has a glycosylation site. 
     
     
         47 . The polynucleotide of  claim 46 , wherein the encoded E38N IFNG polypeptide variant has glycosylation sites at N25, N38 and N97. 
     
     
         48 . The polynucleotide of  claim 36 , wherein the encoded E38N IFNG polypeptide variant comprises an introduced cysteine residue, wherein the introduced cysteine residue is introduced at a residue position that corresponds to a residue position in wild-type human IFNG of SEQ ID NO: 2 that has at least 25% of its side chain surface exposed to solvent. 
     
     
         49 . The polynucleotide of  claim 48 , wherein an introduced cysteine residue of said encoded E38N IFNG polypeptide variant is introduced by substitution at a residue position selected from the group consisting of P3C, K6C, N10C, K13C, N16C, D21C, N25C, G26C, G31C, K34C, K37C, E38C, E39C, K55C, K58C, N59C, D62C, Q64C, S65C, K68C, E71C, E75C, N83C, S84C, K86C, K87C, K94C, N97C, S99C, T101C, D102C, L103C and N104C, relative to the residue positions of wild-type human IFNG of SEQ ID NO: 2. 
     
     
         50 . The polynucleotide of  claim 49 , wherein the introduced cysteine residue of said encoded E38N IFNG polypeptide variant is introduced by substitution at a residue position selected from the group consisting of N25C and N97C. 
     
     
         51 . The polynucleotide of  claim 49 , wherein the introduced cysteine residue of said encoded E38N IFNG polypeptide variant is an N16C substitution. 
     
     
         52 . The polynucleotide of  claim 49 , wherein the introduced cysteine residue of said encoded E38N IFNG polypeptide variant is an N59C substitution. 
     
     
         53 . An expression vector comprising the polynucleotide of  claim 36 . 
     
     
         54 . A host cell transformed with the expression vector of  claim 53 . 
     
     
         55 . The host cell of  claim 54  wherein the host cell is a transformed CHO cell.