Manufacturing and Shelf Stability of Reconstituted High-density Lipoprotein for Infusion Therapy

Although high-density lipoprotein (HDL) infusion therapy now seems to be a promising therapy for the treatment of several diseases, including atherosclerosis, myocardial infarction, sepsis, and diabetes, there is a real need for more information on the production, quality, and safety of reconstituted HDL (rHDL). In this context, we described a production-scale preparation of rHDL and examined the reproducibility of the process and product, and its shelf stability over a 24-month period. Apolipoprotein A-I (apoA-I) was isolated from precipitates Ⅳ (by plasma fractionation) using polyethylene glycol, ethanol, pH precipitation, and ion-exchange chromatography. This manufacturing process included 3 virus-elimination steps consisting of ethanol precipitation, pasteurization, and nano-filtration. HDLs were then reconstituted through cholate dialysis using soybean phosphatidylcholine. The product released data from 3 separate rHDL production were adequate to fulfill the required specification and admitted range. Following lyophilization, the products were stable in the presence of sucrose for at least 24 months. Redissolved rHDLs were disc-shaped and had sizes ranging from 10 to 20 nm. Studies on structure-function relationships provided evidence that these rHDLs could be used as potential therapeutic agents for acute coronary syndrome and inflammatory diseases.