High-yields and extended serum half-life of human interferon alpha2b expressed in tobacco cells as arabinogalactan-protein fusions.
Therapeutic proteins like human interferon alpha2 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 alpha2b (IFNalpha2) in high yields and with increased serum half-life when expressed as arabinogalactan-protein (AGP) chimeras in cultured tobacco cells. Thus IFNalpha2-AGPs targeted for secretion typically gave 350-1400-fold greater secreted yields than the non-glycosylated IFNalpha2 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-IFNalpha2 chimeras showed up to a 13-fold increased in vivo serum half-life while the biological activity remained similar to native IFNalpha2. The use of arabinogalactan glycomodules may provide a general approach to the enhanced production of therapeutic proteins by plants.
Xu
Jianfeng
J
Department of Chemistry and Biochemistry, Biochemistry Research Facility, 350 West State Street, Ohio University, Athens, OH 45701, USA.
Tan
Li
L
Goodrum
Kenneth J
KJ
Kieliszewski
Marcia J
MJ
D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods; D024861:Genetic Enhancement / Q000379:methods; D006207:Half-Life; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000097:blood* / Q000737:chemistry / Q000235:genetics; D008657:Metabolic Clearance Rate; D009088:Mucoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000097:blood / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*
D000077190:Interferon alpha-2; D016898:Interferon-alpha; D009088:Mucoproteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C006619:arabinogalactan proteins