title: Transgenic expression and recovery of biologically active recombinant human insulin from Arabidopsis thaliana seeds.
abstract: 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, DesB(30)-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.
authors.affiliation: SemBioSys Genetics Inc, 110, 2985-23 Ave NE, Calgary, AB, T1Y 7L3, Canada.
authors.forename: Cory L
authors.initials: CL
authors.lastname: Nykiforuk
authors.forename: Joseph G
authors.initials: JG
authors.lastname: Boothe
authors.forename: Elizabeth W
authors.initials: EW
authors.lastname: Murray
authors.forename: Richard G
authors.initials: RG
authors.lastname: Keon
authors.forename: H Joseph
authors.initials: HJ
authors.lastname: Goren
authors.forename: Nancy A
authors.initials: NA
authors.lastname: Markley
authors.forename: Maurice M
authors.initials: MM
authors.lastname: Moloney
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000737:chemistry / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D005818:Genetic Engineering; D006801:Humans; D007328:Insulin / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D030821:Plants, Genetically Modified / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012639:Seeds / Q000737:chemistry / Q000235:genetics*; D014170:Transformation, Genetic; D014357:Trypsin / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D007328:Insulin; D011993:Recombinant Fusion Proteins; C079353:oleosin protein, Arabidopsis; D014357:Trypsin