| About the journal | | | Subscriptions | | | Authors | | | Users | | | Librarians | | | FAQs |
 |
Biotechnology and Applied Biochemistry (1998) 28, (85–94) (Printed in Great Britain)
Chromatographic removal and heat inactivation of hepatitis A virus during manufacture of human albumin
Wayne L. Adcock*1, Andrew MacGregor*, Jeff R. Davies*, Meghan Hattarki*, David A. Anderson† and Neil H. Goss*
*Research and Development, CSL Limited, Bioplasma Division, 189-209 Camp Road, Broadmeadows, Victoria 3047, Australia, and †Macfarlane Burnet Centre for Medical Research, Yarra Bend Road, Fairfield, Victoria 3078, Australia
Abbreviations used: EIA, enzyme immunoassay; HAV, hepatitis A virus; HETP, height equivalent to a theoretical plate; HBV, hepatitis B virus; HCV, hepatitis C virus; MEM, minimal essential medium; RIFA, radioimmunofocus assay; RIFU, radioimmunofocus units; TCID50, tissue culture infectious dose; VAM, virus assay medium; log10, viral loads viral log reduction factors are expressed as log10 of these values and are indicated as n log10. 1 To whom correspondence should be addressed. 
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.
Received 5 December 1997/3 April 1998; accepted 3 April 1998
Portland Press Ltd © 1998 |
Bookmark with:
 
|
PDF
My BAB toolbox
