CHROMATOGRAPHIC REMOVAL AND HEAT INACTIVATION OF HEPATITIS-A VIRUS DURING MANUFACTURE OF HUMAN ALBUMIN

CSL Limited, an Australian biopharmaceutical company, has recently con verted its method of manufacture for human albumin from a traditional Cohn-ethanol fractionation method to a method employing chromatographi c techniques. Studies were undertaken to determine the efficiency of t he chromatographic and pasteurization steps used in the manufacture of Albumex(R) (CSL's trade name for albumin) in removing and inactivatin g the potential viral contaminant, hepatitis A virus (HAV). The manufa cturing process for Albumex(R) includes three chromatographic steps, t wo of which are ion-exchange steps (DEAE-Sepharose(R) Fast Flow and CM -Sepharose(R) Fast Flow) and the third is a gel-filtration step (Sepha cryl(R) S200 HR). The final stage of the Albumex(R) process involves a bulk pasteurization step where product is held at 60 degrees C for IO h, HAV partitioning experiments on the DEAE-Sepharose(R) FF and CM-Se pharose(R) FF ion-exchange and Sephacryl(R) S200 HR gel-filtration col umns were performed with scaled-down models of the production-scale ch romatographic Albumex(R) process, Production samples collected before each of the chromatographic steps were spiked with HAV and processed t hrough each of the scaled-down chromatographic columns, Samples collec ted during processing were assayed and the log,, reduction factors cal culated. Inactivation kinetics of HAV were examined during the pasteur ization of Albumex(R) 5 and 20 [5% and 20% (w/v) albumin solutions] he ld at 60 OC for IO h. Log,, reductions for HAV through the DEAE-Sephar ose(R) FF, CM-Sepharose(R) FF and Sephacryl(R) S200 HR chromatographic columns were 5.3, 1.5 and 4.2 respectively, whereas a 4.4 and a great er than 3.9 log(10) reduction in HAV in Albumex(R) 5 and 20 respective ly were achieved during pasteurization.