PURIFICATION OF RECOMBINANT HUMAN SERUM ALBUMIN FROM THE FERMENTATION BROTH OF Pichia Pastoris GS115/ALBUMIN

Intensive studies on the process of fermentation and purification were conducted in this thesis. During the fermentation, the producing stain Pichia pastoris GS 115/Albumin was induced after 96 hours processing, then the concentration of rI-ISA in the fermentation liquid reached 3.lOg/l. The fermentation liquid was centrifuged at 3,000 r/min for 10 minutes, next the supernatant was heat-treated at 60C for 8 hours and centrifuged to get rid of the sediment. The heat-treated solution was treated with an ultrafiltration membrane having a molecular weight exclusive limit of 30,000, then adding amino sulphate to a final concentration of 80% to precipitate rI-ISA. The precipitated rHSA was separated from the supernatant fluid by centrifugation, redissolved with di-distilled water. The albumin solution obtained in the above processing was applied to a column packed with Phenyl-Sepharose RTM, then the albumin eluted from the column was applied to a column packed with DEAE-Sephadex RTM. The linear elution was carried out and rHSA could be eluted down after half of the linear. The rHSA solution obtained after the purification procedure by ion-exchange chromatography was analyzed by means of HPLC. As shown in the result, the purified rHSA preparation was found as a single peak of HSA monomer. The present technological process is simple and convenient and rHSA thus obtained may be hopeful of passing through the standard of ChP.