Purification of Human Serum Albumin from Plasma with the Combination of Hydrophobic Interaction Chromatography and Cold Ethanol Precipitation
疏水层析结合冷乙醇沉淀纯化人血清白蛋白

Human serum albumin (HSA) has been used to treat a number of diseases with high dosage, so extremely high purity is required in large scale production. HSA from plasma has long been produced by cold ethanol precipitation with purity about 96%, and final recovery 80%. With the development of modern chromatography, its application in purification of HSA has been increasingly studied in the last few years. In this paper, a new process was developed for purification of human serum albumin from plasma. The process features a combination of hydrophobic interaction chromatography and cold ethanol precipitation, hoping to get the product with higher purity, higher recovery and shorter production cycle. The plasma was first treated with cold ethanol in a similar way to the conventional method to eliminate possible virus contamination. The supernatant after desalting and de-ethanol was put through a cation exchange chromatographic column containing CM-Sepharose FF. The operation was in a flow-through mode in which non-albumin protein fractions were retained by the column and albumin fraction passed through. In this way, a fast separation was achieved. Purification of albumin from the flow-through fraction was performed with hydrophobic interaction chromatography which binds albumin tightly. Trace impurities were flushed away from the column. High purity of 99% was obtained as one band in SDS-PAGE with silver staining. The final recovery of HSA was 81.2%. Compared with the traditional process of cold ethanol precipitation, the chromatographic purification can be operated under room temperature, and is faster and more efficient.