Purification and characterization of liposomes encapsulating hemoglobin as potential blood substitutes

In view of the desirability to increase the survival time of the liposome-based artificial red blood cells in vivo, the variables influencing optimum hemoglobin capture and preservation for the bovine hemoglobin-loaded liposomes (LEHb) are investigated. In order to predict the in vivo response, the necessary experiments for the in vitro system characterization have been carried out. The liposomes are prepared by the Reverse Phase Evaporation technique and then purified using a Sepharose 4B column. The purified LEHbs display a unimodal size distribution in the submicron range with a volume average diameter of 0.115 microns and a particle count of 1.25* 10(15) per ml of suspension. Analysis of the lipi/Hb content of the liposomes reveals that the variations in the ratio of Hb encapsulated to lipid entrapped (Hb/L)f as a function of the initial Hb concentration ([Hb]o) is insignificant compared to the net augmentation of (Hb/L)f as a function of the increasing initial lipid to Hb loading ([L]o). Meanwhile high [Hb]o s are necessary for the preservation of oxyhemoglobin.