NR AXZL
AU You,B.; Mornac,S.; Arrabal,S.; Kimmel-Jehan,C.; Flan,B.
TI Investigation of Prion Decontamination from Different Chromatographic Gels by Sodium Hydroxyde
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Natural and Experimental Strains P02.36
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB Sodium hydroxide (NaOH) is a one of the recommended treatments for the decontamination of transmissible spongiform encephalopathies (TSE) agents. The World Health Organization recommends the use of 1N NaOH for 1 hour at room temperature for a complete inactivation of these agents. Such recommendations are based on the results of inactivation experiments that were performed in liquid phase on crude brain homogenates, and that did not include kinetics of inactivation. Few studies are available regarding the efficiency of NaOH treatments on different kind of surfaces and matrixes, including the chromatography gels which are used in the purification of plasma derived products. We have assesed the efficiency of prion decontamination by NaOH on two ion-exchange chromatography gels by using a screening method on a batch format (DEAE Toyopearl and CM-LS Tris-acryl gels). A bead suspension was loaded in a tube and put in contact with crude brain homogenate (0.2% w/v - 263K hamster strain of scrapie). The unbound fraction was discarded and the remaining beads were treated with different concentrations of NaOH under severall conditions of time and temperature. The beads were rinsed and submitted to elution in NaCl 2M. The amount of PrP-sc was quantified by Western blot in both the elution solution and remaining beads suspension. The reduction factors (RF) were calculted relative to the level of PrP-sc detected in spiked beads that were treated with water instead of NaOH. The results showed efficient PrP-sc inactivation on a DEAE Toyopearl gel with 0.2M NaOH (RF > 4 log10 for the elution solution and RF > 2.5 log10 for the bead suspension) in a short time (18 minutes) at room temperature. Similarly, efficient PrP-sc inactivation was observed on a CM-LS Tris-acryl gel treated with 0.2M NaOH (RF > 3.5 log10 for the elution solution and RF > 2.5 log10 for the bead suspension) after 60 minutes at 4°. Taken together, these results show that NaOH treatments are efficient in inactivating PrP-sc on DEAE toyopearl and CM-LS Tris-acryl chromatography gels, with no PrP-sc detected on treated beads. These experiments will be extented to other gels and should be completed by an infectivity assay. In conclusion, the chromatography gels treatment by NaOH, that is already applied in industrial conditions, contribute to the safety of plasma products regarding the theoretical risk of vCJD trasmission.
AD B. You, S. Mornac, S. Arrabal, C. Kimmel-Jehan, B. Flan, Laboratoire du Fractionnement et des Biotechnologies, Unité de Sécurisation Biologique, France
SP englisch
PO Schottland