Prion-removal capacity of chromatographic and ethanol precipitation steps used in the production of albumin and immunoglobulins
Article first published online: 8 NOV 2006
Volume 91, Issue 4, pages 292–300, November 2006
How to Cite
Thyer, J., Unal, A., Thomas, P., Eaton, B., Bhashyam, R., Ortenburg, J., Uren, E., Middleton, D., Selleck, P. and Maher, D. (2006), Prion-removal capacity of chromatographic and ethanol precipitation steps used in the production of albumin and immunoglobulins. Vox Sanguinis, 91: 292–300. doi: 10.1111/j.1423-0410.2006.00829.x
- Issue published online: 20 NOV 2006
- Article first published online: 8 NOV 2006
- Received: 28 February 2006, revised 27 June 2006, accepted 3 July 2006
- transmissible spongiform encephalopathy;
- Western blot
Background and Objectives Although there is no epidemiological evidence to suggest that classical Creutzfeldt–Jakob disease (CJD) is transmitted through blood or blood products, the variant form (vCJD) has been implicated in transmission via packed red blood cells. The potential threat of the infectious agent contaminating plasma pools has led to manufacturing processes being examined for capacity to remove prions. The objective of these studies was to examine the prion-removal potential of the chromatographic purification and ethanol precipitation steps used to fractionate immunoglobulins and albumin from human plasma.
Materials and Methods Western blot assay was used to examine the partitioning of proteinase K-resistant scrapie prion protein (PrPsc) over DEAE Sepharose, CM Sepharose and Macro-Prep High Q chromatographic columns, utilizing microsomal scrapie 263K spiked into each scaled down feedstream and assayed after each chromatographic step. In further studies, bioassay in C57 black mice was used and spikes of 10 000 g clarified brain homogenate of scrapie ME7 were added to feedstreams before sequences of scaled down chromatographic or Cohn fractionation process steps.
Results The microsomal spiking study with Western blot detection demonstrated substantial partitioning of PrPsc away from the target proteins in all ion exchange chromatographic steps examined. The log10 reduction factors (LRF) across DEAE Sepharose and CM Sepharose columns for albumin were ≥ 4.0 and ≥ 3.0 respectively. The reductions across DEAE Sepharose and Macro-Prep High Q for intravenous immunoglobulin were 3.3 and ≥ 4.1 respectively. Bioassay demonstrated LRFs of ≥ 5.6 across the combination of DEAE Sepharose and CM Sepharose columns in the albumin process and ≥ 5.4 across the combination of DEAE Sepharose and Macro-Prep High Q columns in the intravenous immunoglobulin process. Bioassay studies also demonstrated a LRF of ≥ 5.6 for immunoglobulin produced by Cohn fractionation.
Conclusions Using rodent-adapted scrapie as a model, the studies indicated that ion exchange chromatography, as well as Cohn immunoglobulin fractionation have the potential to effectively reduce the load of TSE agents should they be present in plasma pools.
Table of Contents Ion exchange columns used for production of human albumin and immunoglobulins, as well as Cohn immunoglobulin fractionation, effectively reduce the load of TSE agents should they be present in plasma pools.