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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Chuang, Tzu-ying Spooner, Eric Popp, Maximilian W. Ploegh, Hidde L. Dougan, Stephanie K. |
| Description | Author Affiliation: Popp MW ( Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, MA 02142, USA.); |
| Abstract | Recombinant protein therapeutics often suffer from short circulating half-life and poor stability, necessitating multiple injections and resulting in limited shelf-life. Conjugation to polyethylene glycol chains (PEG) extends the circulatory half-life of many proteins, but the methods for attachment often lack specificity, resulting in loss of biological activity. Using four-helix bundle cytokines as an example, we present a general platform that uses sortase-mediated transpeptidation to facilitate site-specific attachment of PEG to extend cytokine half-life with full retention of biological activity. Covalently joining the N and C termini of proteins to obtain circular polypeptides, again executed using sortase, increases thermal stability. We combined both PEGylation and circularization by exploiting two distinct sortase enzymes and the use of a molecular suture that allows both site-specific PEGylation and covalent closure. The method developed is general, uses a set of easily accessible reagents, and should be applicable to a wide variety of proteins, provided that their termini are not involved in receptor binding or function. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 8 |
| Volume Number | 108 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2011-03-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Aminoacyltransferases Chemistry Bacterial Proteins Cysteine Endopeptidases Polyethylene Glycols Protein Stability Administration & Dosage Metabolism Animals Catalysis Cyclization Cytokines Half-Life Mice Streptococcus Pyogenes Research Support, N.I.H., Extramural Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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