Glycoprotein synthesis

Abstract

Methods for making glycoproteins, both in vitro and in vivo, are provided. One method involves incorporating an unnatural amino acid into a protein and attaching one or more saccharide moieties to the unnatural amino acid. Another method involves incorporating an unnatural amino acid that includes a saccharide moiety into a protein. Proteins made by both methods can be further modified with additional sugars.

1 - 57 . (canceled) 
     
     
         58 . A composition comprising an antibody that comprises an unnatural amino acid, wherein the composition additionally comprises a recombinant glycosylase that glycosylates the unnatural amino acid. 
     
     
         59 . The composition of  claim 58 , wherein the antibody is a polyclonal, monoclonal, chimeric, or single chain antibody. 
     
     
         60 . The composition of  claim 58 , wherein the antibody is a fragment of a full-length immunoglobulin. 
     
     
         61 . The composition of  claim 58 , wherein the composition comprises a translation system, cell or cell lysate that comprises the antibody and the recombinant glycosylase. 
     
     
         62 . The composition of  claim 61 , wherein the translation system, cell, or cell lysate comprises a recombinant nucleic acid that comprises a selector codon and that encodes at least one polypeptide chain of the antibody, an orthogonal tRNA (OtRNA) that recognizes the selector codon, and an orthogonal aminoacyl tRNA synthetase (ORS) that aminoacylates the OtRNA with the unnatural amino acid, wherein the translation system, cell or cell lysate incorporates the unnatural amino acid into the antibody in response to the selector codon.