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. Apolipoprotein A-I (apoA-I was isolated from precipitates obtained by cold ethanol fractionation of human plasma. This process includes several steps for virus removal and virus inactivation, among them pasteurization. Reconstitution of lipoprotein particles was performed by cholate dialysis 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 protein as well as the low concentration of free lipid. The product was stabilized by lyophilization in the presence of sucrose. In vitro studies show potential effects it the prevention of gram-negative septic shock and in the inhibition of atherosclerosis.
ZLB Central Laboratory, Blood Transfusion Service SRC, Bern, Switzerland.
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Lerch
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Förtsch
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Hodler
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Bolli
D016632:Apolipoprotein A-I / Q000302:isolation & purification; D002850:Chromatography, Gel; D004789:Enzyme Activation; D006801:Humans; D008070:Lipopolysaccharides / Q000378:metabolism; D008075:Lipoproteins, HDL / Q000737:chemistry*; D008970:Molecular Weight; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000378:metabolism; D010713:Phosphatidylcholines
D016632:Apolipoprotein A-I; D008070:Lipopolysaccharides; D008075:Lipoproteins, HDL; D010713:Phosphatidylcholines; D007862:Phosphatidylcholine-Sterol O-Acyltransferase