A Highly Bioactive Lys-Deficient IFN Leads to a Site-Specific Di-PEGylated IFN with Equivalent Bioactivity to That of Unmodified IFN-α2b
| Autor: | Takehiro Suzuki, Michinori Kohara, Takashi Imada, Naoshi Dohmae, Yo Ichi Tagawa, Koji Moriya, Mitsuhiro Hitotsuyanagi, Naoto Inukai, Masahiko Uchiyama, Daisuke Kiga, Shotaro Ayukawa, Akiko Masuda, Masayuki Yamamura |
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| Rok vydání: | 2018 |
| Předmět: |
Biomedical Engineering
Polyethylene glycol Interferon alpha-2 Antiviral Agents Biochemistry Genetics and Molecular Biology (miscellaneous) Polyethylene Glycols Mice chemistry.chemical_compound Pharmacokinetics In vivo Interferon medicine Animals Humans Protein Stability Chemistry Lysine Interferon-alpha General Medicine Directed evolution Recombinant Proteins In vitro A-site Biochemistry Mutagenesis Site-Directed PEGylation Directed Molecular Evolution medicine.drug |
| Zdroj: | ACS Synthetic Biology. 7:2537-2546 |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.8b00188 |
| Popis: | Although conjugation with polyethylene glycol (PEGylation) improves the pharmacokinetics of therapeutic proteins, it drastically decreases their bioactivity. Site-specific PEGylation counters the reduction in bioactivity, but developing PEGylated proteins with equivalent bioactivity to that of their unmodified counterparts remains challenging. This study aimed to generate PEGylated proteins with equivalent bioactivity to that of unmodified counterparts. Using interferon (IFN) as a model protein, a highly bioactive Lys-deficient protein variant generated using our unique directed evolution methods enables the design of a site-specific di-PEGylated protein. Antiviral activity of our di-PEGylated IFN was similar to that of unmodified IFN-α2b. The di-PEGylated IFN exhibited 3.0-fold greater antiviral activity than that of a commercial PEGylated IFN. Moreover, our di-PEGylated IFN showed higher in vitro and in vivo stability than those of unmodified IFN-α2b. Hence, we propose that highly bioactive Lys-deficient proteins solve the limitation of conventional PEGylation with respect to the reduction in bioactivity of PEGylated proteins. |
| Databáze: | OpenAIRE |
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