Isolation and purification of albumin from human plasma by direct and combined approach of ion-exchange chromatography and comparison of the final products quality obtained by both methods

Due to multiple roles of albumin in the body, injection of its medicinal product as one of the therapeutic or management strategies under conditions such as severe bleeding, burns, liver failure, and neonatal hemolytic diseases is on the physicians' agenda. Considering that albumin is the most abundant plasma protein, designing an appropriate method to purify it is highly important. There are several methods such as human plasma fractionation, chromatographic, or Salting-out methods for the isolation and purification of the human albumin. The present study investigates a direct and combined ion-exchange chromatography approach for purification of albumin from human plasma and compares the quality of the final products obtained by both ion-exchange chromatographic methods.

This study was carried out from January 2019 to October 2019 at the Blood Transfusion Research Center, High institute for research and education in transfusion medicine, affiliated with the Iranian Blood Transfusion Organization. For this study, 10 human plasma bags were randomly collected. After thawing, all 10 human plasma bags were pooled, and in order to separate cryo paste, it was centrifuged at 4000 g for 10 minutes at the temperature of 1 Centigrade degree. Then the obtained cryo poor plasma was used to purify the albumin protein by direct and combined methods of ionexchange chromatography. The purity of the final products was compared by cellulose acetate electrophoresis and SDS-PAGE tests. The sample obtained by the combined approach was pasteurized and HPLC analysis was performed to investigate any polymer aggregates.

In contrast to the direct method, the final product obtained by combined ionexchange chromatography had a good purity by the average of about 95% and the amount of polymer was estimated to be less than 5% by HPLC analysis (P<0.05).

By diluting the plasma and subsequently reducing the ionic strength, albumin can be separated from human plasma with a high degree of purity only by two steps of ion-exchange chromatography