Transgenic expression and recovery of biologically active recombinant human insulin from Arabidopsis thaliana seeds

Summary The increased incidence of diabetes, coupled with the introduction of alternative delivery methods that rely on higher doses, is expected to result in a substantial escalation in the demand for affordable insulin in the future. Limitations in the capacity and economics of production will mak...

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Published in:Plant biotechnology journal 2006-01, Vol.4 (1), p.77-85
Main Authors: Nykiforuk, Cory L., Boothe, Joseph G., Murray, Elizabeth W., Keon, Richard G., Goren, H. Joseph, Markley, Nancy A., Moloney, Maurice M.
Format: Article
Language:English
Subjects:
Animals
Arabidopsis
Arabidopsis - chemistry
Arabidopsis - genetics
Arabidopsis Proteins - genetics
Arabidopsis thaliana
Cell Line, Tumor
diabetes
Genetic Engineering
Humans
insulin
Insulin - genetics
Insulin - isolation & purification
Insulin - metabolism
Male
Mice
Mice, Inbred C57BL
oleosin
Phosphorylation
Plants, Genetically Modified - genetics
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - isolation & purification
Recombinant Fusion Proteins - metabolism
recombinant protein
Seeds - chemistry
Seeds - genetics
Transformation, Genetic
Trypsin - metabolism
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Summary:Summary The increased incidence of diabetes, coupled with the introduction of alternative delivery methods that rely on higher doses, is expected to result in a substantial escalation in the demand for affordable insulin in the future. Limitations in the capacity and economics of production will make it difficult for current manufacturing technologies to meet this demand. We have developed a novel expression and recovery technology for the economical manufacture of biopharmaceuticals from oilseeds. Using this technology, recombinant human precursor insulin was expressed in transgenic plants. Plant‐derived insulin accumulates to significant levels in transgenic seed (0.13% total seed protein) and can be enzymatically treated in vitro to generate a product with a mass identical to that of the predicted product, DesB30‐insulin. The biological activity of this product in vivo and in vitro was demonstrated using an insulin tolerance test in mice and phosphorylation assay performed in a mammalian cell culture system, respectively.
ISSN:1467-7644
1467-7652
DOI:10.1111/j.1467-7652.2005.00159.x