PRODUCTION AND CHARACTERIZATION OF A RECONSTITUTED HIGH-DENSITY-LIPOPROTEIN FOR THERAPEUTIC APPLICATIONS

A method is described for the large scale preparation of reconstituted high density lipoproteins (rHDL) suitable for therapeutic use. Apolip oprotein A-I (apoA-I) was isolated from precipitates obtained by cold ethanol fractionation of human plasma. This process includes several s teps for virus removal and virus inactivation, among them pasteurizati on. Reconstitution of lipoprotein particles was performed by cholate d ialysis using soybean phosphatidylcholine as the lipid source. An apoA -I:lipid ratio of 1:150 (mol:mol) was obtained. Redissolved rHDLs were disc-shaped particles resembling nascent HDL, as assessed by electron microscopy. The method was optimized for low content of free apoA-I p rotein as well as the low concentration of free lipid. The product was stabilized by lyophilization in the presence of sucrose. In vitro stu dies show potential effects in the prevention of gram-negative septic shock and in the inhibition of atherosclerosis.