Lipoprotein complexes and manufacturing and uses thereof

A method of producing a formulation comprising Apolipoprotein A-I (“ApoA-I”) protein complexed with lipids, comprising:

• (a) culturing in a culture medium a mammalian host cell engineered to express an ApoA-I protein comprising:
• (i) a signal sequence;
• (ii) a propeptide sequence;
• and
• (iii) an amino acid sequence comprising positions 25 to 267 of SEQ ID NO:1,
• wherein the mammalian host cell comprises a nucleic acid encoding the ApoA-I protein operably linked to a constitutive promoter;
• (b) recovering ApoA-I protein from the culture medium;
• (c) purifying the recovered ApoA-I protein;
• and
• (d) complexing the purified ApoA-I with sphingomyelin and 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (“DPPG”) or a salt thereof to yield a composition comprising a population of lipoprotein complexes, wherein the sphingomyelin and the DPPG or the salt thereof are present in the composition in a 97:3 weight:weight ratio and the ApoA-I, the sphingomyelin and the DPPG or the salt thereof are present in the composition in a 1:2.7 ratio of weight ApoA-I:weight sphingomyelin and DPPG or salt thereof.