TY - JOUR
T1 - A Highly Bioactive Lys-Deficient IFN Leads to a Site-Specific Di-PEGylated IFN with Equivalent Bioactivity to That of Unmodified IFN-α2b
AU - Imada, Takashi
AU - Moriya, Koji
AU - Uchiyama, Masahiko
AU - Inukai, Naoto
AU - Hitotsuyanagi, Mitsuhiro
AU - Masuda, Akiko
AU - Suzuki, Takehiro
AU - Ayukawa, Shotaro
AU - Tagawa, Yo Ichi
AU - Dohmae, Naoshi
AU - Kohara, Michinori
AU - Yamamura, Masayuki
AU - Kiga, Daisuke
N1 - Funding Information:
This research was partially supported by a Gamma Project Grant for T.I. from the Education Academy of Computational Life Science of the Tokyo Institute of Technology a Waseda University Grant for Special Research Projects (Special Research 2017B-226) for D.K. from Waseda University, a Grant-in-Aid for Scientific Research (KAKENHI, 15KT0148) for S.A. from the Ministry of Education Science and Culture of Japan and a Grant for Industrial Technology Research for D.K from New Energy and Technology Developmental Organization (Industrial Technology Research Grant Program 05A02512d).
Funding Information:
We thank Kanako Azami for technical support, Hiroshi Nakayama, Seketsu Fukuzawa, and Koichiro Kodama for the analysis of the simplified genetic code, and Kazuyoshi Mihara, Hiroshi Tsutsumi, and Department of Technical Support and Promotion of Shared Research Apparatus for the analysis of CD in the study. This research was partially supported by a Gamma Project Grant for T.I. from the Education Academy of Computational Life Science of the Tokyo Institute of Technology, a Waseda University Grant for Special Research Projects (Special Research 2017B-226) for D.K. from Waseda University, a Grant-in-Aid for Scientific Research (KAKENHI, 15KT0148) for S.A. from the Ministry of Education, Science and Culture of Japan and a Grant for Industrial Technology Research for D.K from New Energy and Technology Developmental Organization (Industrial Technology Research Grant Program 05A02512d).
Publisher Copyright:
© 2018 American Chemical Society.
PY - 2018/11/16
Y1 - 2018/11/16
N2 - 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.
AB - 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.
KW - Lys-deficient proteins
KW - directed evolution
KW - genetic code engineering
KW - interferon
KW - site-specific di-PEGylation
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U2 - 10.1021/acssynbio.8b00188
DO - 10.1021/acssynbio.8b00188
M3 - Article
C2 - 30277749
AN - SCOPUS:85056135406
SN - 2161-5063
VL - 7
SP - 2537
EP - 2546
JO - ACS Synthetic Biology
JF - ACS Synthetic Biology
IS - 11
ER -