Site-specific PEGylation of proteins containing unnatural amino acids

We report a generally applicable PEGylation methodology based on the site-specific incorporation ofpara-azidophenylalanine into proteins in yeast

Alexander Deiters

2004

Scholarcy highlights

  • Despite the importance of proteins as therapeutic agents, they have a number of disadvantages in comparison to small molecule drugs, including poor oral bioavailability and in many cases short serum halflives
  • The lack of selectivity and positional control in the attachment of PEG chains can lead to significant losses in biological activity and possibly enhanced immunogenicity of the conjugated protein
  • We report a novel method that allows for the completely site specific and irreversible attachment of PEG chains to proteins based on unnatural amino acid mutagenesis
  • It was shown that amino acids with alkenyl, azido, and keto side chains can be used to efficiently couple sugars, biotin, and fluorescent pHruoibsegsentootceyinclso."ad-d' 3itiBonecl 4au-1s6eotfhaen copper-mediated azide and an alkyne is orthogonal to all functional groups found in proteins and forms a stable triazole linkage, we investigated this reaction for the selective PEGylation of proteins
  • Based on the crystal structure of SOD, 18 the surface exposed residue Trp33 was selected as the PEGylation site, and the corresponding codon was mutated to TAG
  • For comparison wild type SOD was pegylated with commonly used N-hydroxysuccinimide esters 2 and 3, leading to a multitude of conjugated proteins as visualized by SOS-PAGE. 19
  • We have developed a novel PEGylation technology, which allows the site specific and irreversible attachment of a single PEG molecule to a protein

Need more features? Save interactive summary cards to your Scholarcy Library.