In-plant production of dimeric and/or oligomeric (comprising three or more units) forms of human apo a-1 protein muteins

A genetically transformed plant with one or more expression vectors comprising a seed-specific expression cassette and a nucleotide sequence encoding a mutein of human apolipoprotein Apo A-1, expressing in the seed's storage tissue one or more of said muteins, and forming dimers and/or oligomers, comprising three or more monomers, in an essentially dimeric and/or oligomeric form, of said muteins characterised in that they show in their dimeric and/or oligomeric forms, biological activities equal or higher of the activities shown by apolipoprotein Apo A-1 Milano in its dimeric form in the following tests:

• a′. measurement of the reverse cholesterol transport capacity evaluated by calculation of the association kinetics of equal quantities of said dimer or of said oligomer, and of the said dimer of Apo A-1 Milano with dimyristoylphosphatidylcholine (DMPC);
• b′. measurement of the plasma half life of said dimer or of said oligomer by a plasma half life test in comparison to a calibration curve obtained performing the same test with the purified Apo A-1 Milano dimer.

A seed comprising in its storage organs one or more muteins of the human apolipoprotein Apo A-1 capable of forming dimers and/or oligomers, comprising three or more monomers, in an essentially dimeric and/or oligomeric form;

• said muteins characterised in that they show in their dimeric and/or oligomeric forms, biological activities equal or higher of the activities shown by apolipoprotein Apo A-1 Milano in its dimeric form in the following tests:
• a′. measurement of the reverse cholesterol transport capacity evaluated by calculation of the association kinetics of equal quantities of said dimer or of said oligomer, and of the said dimer of Apo A-1 Milano with dimyristoylphosphatidylcholine (DMPC);
• b′. measurement of the plasma half life of said dimer or of said oligomer by a plasma half life test in comparison to a calibration curve obtained performing the same test with the purified Apo A-1 Milano dimer.