Polypeptides With Enhanced Anti-Inflammatory And Decreased Cytotoxic Properties And Relating Methods

Abstract

The invention provides a polypeptide containing at least one IgG Fc region, wherein said at least one IgG Fc region is glycosylated with at least one galactose moiety connected to a respective terminal sialic acid moiety by a α 2,6 linkage, and wherein said polypeptide having a higher anti-inflammatory activity as compared to an unpurified antibody.

1 .- 36 . (canceled) 
     
     
         37 . A method for preparing a pharmaceutical preparation comprising a recombinant IgG antibody having increased anti-inflammatory activity, the method comprising:
 expressing, in a host cell, a recombinant IgG antibody;   subjecting the expressed recombinant IgG antibody to one or more purification steps to produce a preparation comprising the recombinant IgG antibody;   treating the composition comprising the recombinant IgG antibody with an alpha-(2,6) sialylatransferase and a donor of sialic acid to increase the fraction of recombinant IgG antibody in the composition having alpha-2,6-linked N-acetylneuraminic acid on N-glycans of the Fc region of the recombinant IgG antibody thereby preparing a composition enriched for sialylated recombinant IgG antibody, wherein the enriched composition has increased anti-inflammatory activity relative to the un-enriched composition, and   processing the composition enriched for sialylated recombinant IgG antibody to produce a pharmaceutical preparation, wherein the processing comprises combining the recombinant IgG antibody with a physiologically acceptable carrier, excipient or stabilizer, thereby preparing a pharmaceutical preparation comprising a recombinant IgG antibody having increased anti-inflammatory activity.   
     
     
         38 . The method of  claim 37 , wherein the recombinant IgG antibody is a monoclonal antibody. 
     
     
         39 . The method of  claim 37 , wherein the recombinant IgG antibody is chimeric antibody. 
     
     
         40 . The method of  claim 37 , wherein the recombinant IgG antibody is a humanized antibody. 
     
     
         41 . The method of  claim 37 , where the composition enriched for sialylated recombinant IgG antibody to affinity chromatography to produce a composition further enriched for sialylated recombinant IgG antibody. 
     
     
         42 . The method of  claim 37 , wherein the host cell is a mammalian cell. 
     
     
         43 . The method of  claim 42 , wherein the mammalian cell line is a CHO cell. 
     
     
         44 . The method of  claim 37 , wherein the purification step comprises ion-exchange chromatography or affinity chromatography. 
     
     
         40 . The method of  claim 37 , wherein the composition enriched for sialylated recombinant IgG antibody exhibits an increased protective effect in a K/B×N serum-induced arthritis mouse model relative to the un-enriched composition. 
     
     
         41 . The method of  claim 37 , wherein the host cell harbors a recombinant sialyltransferase.