Plant expression of cocaine hydrolase-Fc fusion protein for treatment of cocaine abuseReport as inadecuate

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BMC Biotechnology

, 16:72

Protein and enzyme technology


BackgroundA 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.

ResultsCocH3-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.

ConclusionsIt 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.

KeywordsTherapeutic protein Fusion protein Protein production Drug abuse AbbreviationsBChEButyrylcholinesterase

CHOChinese hamster ovary

CNSCentral nervous system

CocHCocaine hydrolase

CocH1Cocaine hydrolase 1

CocH3Cocaine hydrolase 3

CocH3-FcCocaine hydrolase 3 fused with Fc region of human immunoglobulin G1

EREndoplasmic reticulum

Fcfragment crystallizable Fc region of human immunoglobulin G1

HSAHuman serum albumin

IgG1Immunoglobulin G1

pBChEplant-expressed BChE protein

pCocH3plant-expressed CocH3 protein

pCocH3-FcPlant-expressed CocH3-Fc protein

PEG-pCocH3-FcPEGylated pCocH3-Fc

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Author: Guojun Wang - Ting Zhang - Haifeng Huang - Shurong Hou - Xiabin Chen - Fang Zheng - Chang-Guo Zhan


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