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High‐yields and extended serum half‐life of human interferon α2b expressed in tobacco cells as arabinogalactan‐protein fusions
Author(s) -
Xu Jianfeng,
Tan Li,
Goodrum Kenneth J.,
Kieliszewski Marcia J.
Publication year - 2007
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21407
Subject(s) - arabinogalactan , glycoprotein , interferon , fusion protein , in vivo , proteases , biochemistry , biological activity , chemistry , biology , in vitro , virology , polysaccharide , recombinant dna , gene , microbiology and biotechnology , enzyme
Therapeutic proteins like human interferon α2 generally possess short serum half‐lives due to their small size, hence rapid renal clearance, and susceptibility to serum proteases. Chemical derivatization, such as addition of polyethylene glycol (PEG) groups overcomes both problems, but at the expense of greatly decreased bioactivity. We describe a new method that yields biologically potent interferon α2b (IFNα2) in high yields and with increased serum half‐life when expressed as arabinogalactan‐protein (AGP) chimeras in cultured tobacco cells. Thus IFNα2‐AGPs targeted for secretion typically gave 350–1400‐fold greater secreted yields than the non‐glycosylated IFNα2 control. The purified AGP domain itself was not immunogenic when injected into mice and only mildly so when injected as a fusion glycoprotein. Importantly, the AGP‐IFNα2 chimeras showed up to a 13‐fold increased in vivo serum half‐life while the biological activity remained similar to native IFNα2. The use of arabinogalactan glycomodules may provide a general approach to the enhanced production of therapeutic proteins by plants. Biotechnol. Bioeng. 2007; 97: 997–1008. © 2007 Wiley Periodicals, Inc.