title: Expression and recovery of biologically active recombinant Apolipoprotein AI(Milano) from transgenic safflower (Carthamus tinctorius) seeds.
abstract: Apolipoprotein AI Milano (ApoAI(Milano) ) was expressed as a fusion protein in transgenic safflower seeds. High levels of expression corresponding to 7 g of ApoAI(Milano) per kilogram of seed have been identified in a line selected for commercialization. The ApoAI(Milano) fusion protein was extracted from seed using an oilbody-based process and matured in vitro prior to final purification. This yielded a Des-1,2-ApoAI(Milano) product which was confirmed by biochemical characterization including immunoreactivity against ApoAI antibodies, isoelectric point, N-terminal sequencing and electrospray mass spectrometry. Purified Des-1,2-ApoAI(Milano) readily associated with dimyristoylphosphatidylcholine in clearance assays comparable to Human ApoAI. Its biological activity was assessed by cholesterol efflux assays using Des-1,2-ApoAI(Milano) :1-palmitoyl-2-oleoyl phosphatidylcholine complexes in vitro and in vivo. This study has established that high levels of biologically functional ApoAI(Milano) can be produced using a plant-based expression system.
authors.affiliation: SemBioSys Genetics Inc., Calgary, Alberta, Canada. nykiforukc@sembiosys.com
authors.forename: Cory L
authors.initials: CL
authors.lastname: Nykiforuk
authors.forename: Yin
authors.initials: Y
authors.lastname: Shen
authors.forename: Elizabeth W
authors.initials: EW
authors.lastname: Murray
authors.forename: Joseph G
authors.initials: JG
authors.lastname: Boothe
authors.forename: David
authors.initials: D
authors.lastname: Busseuil
authors.forename: Eric
authors.initials: E
authors.lastname: Rhéaume
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Tardif
authors.forename: Alexandra
authors.initials: A
authors.lastname: Reid
authors.forename: Maurice M
authors.initials: MM
authors.lastname: Moloney
mesh_terms: D000818:Animals; D016632:Apolipoprotein A-I / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D031187:Carthamus tinctorius / Q000235:genetics* / Q000378:metabolism; D002784:Cholesterol / Q000097:blood; D050356:Lipid Metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D010713:Phosphatidylcholines / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012639:Seeds / Q000235:genetics* / Q000378:metabolism
chemical_list: D016632:Apolipoprotein A-I; D010713:Phosphatidylcholines; D011993:Recombinant Fusion Proteins; D002784:Cholesterol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine