Chromatographic removal and heat inactivation of hepatitis A virus during manufacture of human albumin

Abstract

CSL Limited, an Australian biopharmaceutical company, has recently converted its method of manufacture for human albumin from a traditional Cohn‐ethanol fractionation method to a method employing chromatographic techniques. Studies were undertaken to determine the efficiency of the chromatographic and pasteurization steps used in the manufacture of Albumex® (CSL's trade name for albumin) in removing and inactivating the potential viral contaminant, hepatitis A virus (HAV). The manufacturing process for Albumex® includes three chromatographic steps, two of which are ion‐exchange steps (DEAE‐ Sepharose® Fast Flow and CM‐Sepharose® Fast Flow) and the third is a gel‐filtration step (Sephacryl® S200 HR). The final stage of the Albumex® process involves a bulk pasteurization step where product is held at 60 °C for 10 h. HAV partitioning experiments on the DEAE‐Sepharose® FF and CM‐Sepharose® FF ion‐ exchange and Sephacryl® S200 HR gel‐filtration columns were performed with scaled‐down models of the production‐scale chromatographic Albumex® process. Production samples collected before each of the chromatographic steps were spiked with HAV and processed through each of the scaled‐down chromatographic columns. Samples collected during processing were assayed and the log10 reduction factors calculated. Inactivation kinetics of HAV were examined during the pasteurization of Albumex® 5 and 20 [5% and 20% (w/v) albumin solutions] held at 60 °C for 10 h. Log10 reductions for HAV through the DEAE‐Sepharose® FF, CM‐ Sepharose® FF and Sephacryl® S200 HR chromatographic columns were 5.3, 1.5 and 4.2 respectively, whereas a 4.4 and a greater than 3.9 log10 reduction in HAV in Albumex® 5 and 20 respectively were achieved during pasteurization.