Recombinant Protein Hydrazides: Application to Site-Specific Protein PEGylation
| Autor: | David E. Anderson, Jennifer Thom, Graham Cotton, Joanne McGregor |
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| Rok vydání: | 2011 |
| Předmět: |
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Molecular Specific protein Biomedical Engineering Pharmaceutical Science Bioengineering Microbial Sensitivity Tests Interferon alpha-2 Protein Engineering Antiviral Agents Polyethylene Glycols law.invention Structure-Activity Relationship Protein sequencing FLAG-tag law Cell Line Tumor Humans Structure–activity relationship Encephalomyocarditis virus Pharmacology Chemistry Organic Chemistry Hydrazones Interferon-alpha Interferon-beta Combinatorial chemistry Recombinant Proteins Biochemistry Cell culture PEGylation Recombinant DNA Interferon beta-1b Biotechnology Chemical Cleavage |
| Zdroj: | Bioconjugate Chemistry. 22:1017-1020 |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/bc2001374 |
| Popis: | Here, we describe a novel method for the site-specific C-terminal PEGylation of recombinant proteins. This general approach exploits chemical cleavage of precursor intein-fusion proteins with hydrazine to directly produce recombinant protein hydrazides. This unique functionality within the protein sequence then facilitates site-specific C-terminal modification by hydrazone-forming ligation reactions. This approach was used to generate folded, site-specifically C-terminal PEGylated IFNalpha2b and IFNbeta1b, which retained excellent antiviral activity, demonstrating the utility of this technology in the PEGylation of therapeutic proteins. As this methodology is straightforward to perform, is compatible with disulfide bonds, and is exclusively selective for the protein C-terminus, it shows great potential as general technology for the site-specific engineering and labeling of recombinant proteins. |
| Databáze: | OpenAIRE |
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