BACKGROUND: A recently reported cocaine hydrolase (CocH3) fused with fragment crystallizable (Fc) region of human immunoglobulin G1, denoted as CocH3-Fc, is known as a promising therapeutic candidate for the treatment of cocaine overdose and addiction. A challenge for practical therapeutic use of this enzyme exists in the large-scale protein production and, therefore, it is interesting to identify a low-cost and feasible, sustainable source of CocH3-Fc production.
RESULTS: CocH3-Fc was transiently expressed in plant Nicotiana benthamiana leaves. The plant-expressed protein, denoted as pCocH3-Fc, was as active as that expressed in mammalian cells both in vitro and in vivo. However, compared to the mammalian-cell expressed CocH3-Fc protein, pCocH3-Fc had a shorter biological half-life, probably due to the lack of protein sialylation in plant. Nevertheless, the in vivo half-life was significantly extended upon the PEGylation of pCocH3-Fc. The Fc fusion did not prolong the biological half-life of the plant-expressed enzyme pCocH3-Fc, but increased the yield of the enzyme expression in the plant under the same experimental conditions.
CONCLUSIONS: It is feasible to express pCocH3-Fc in plants. Further studies on the pCocH3-Fc production in plants should focus on the development of vectors with additional genes/promoters for the complete protein sialylation and for a better yield.
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This work was supported by the National Institutes of Health (NIH) through the NIDA Translational Avant-Garde Award (UH2 DA041115) and R01 grants (R01 DA035552, R01 DA032910, R01 DA013930, and R01 DA025100).
The datasets during and/or analyzed during the current study available from the corresponding author on reasonable request.
Wang, Guojun; Zhang, Ting; Huang, Haifeng; Hou, Shurong; Chen, Xiabin; Zheng, Fang; and Zhan, Chang-Guo, "Plant Expression of Cocaine Hydrolase-Fc Fusion Protein for Treatment of Cocaine Abuse" (2016). Molecular Modeling and Biopharmaceutical Center Faculty Publications. 2.