Expression and recovery of biologically active recombinant Apolipoprotein AI(Milano) from transgenic safflower (Carthamus tinctorius) seeds

Plant Biotechnol J. 2011 Feb;9(2):250-63. doi: 10.1111/j.1467-7652.2010.00546.x.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apolipoprotein A-I / genetics*
  • Apolipoprotein A-I / metabolism
  • Apolipoprotein A-I / pharmacology
  • Carthamus tinctorius / genetics*
  • Carthamus tinctorius / metabolism
  • Cholesterol / blood
  • Lipid Metabolism
  • Mice
  • Mice, Inbred C57BL
  • Phosphatidylcholines / genetics
  • Phosphatidylcholines / metabolism
  • Phosphatidylcholines / pharmacology
  • Plants, Genetically Modified / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Seeds / genetics*
  • Seeds / metabolism


  • Apolipoprotein A-I
  • Phosphatidylcholines
  • Recombinant Fusion Proteins
  • Cholesterol
  • 1-palmitoyl-2-oleoylphosphatidylcholine