Validation of Virus Inactivation by Heat Treatment in the Manufacture of Diaspqun Crosslinksd Hemoglobin

Publisher Website
Google Scholar
Abstract
Diaspirin crosslinked hemoglobin (DCLHb), a hemoglobin based oxygen carrying solution prepared from outdated human blood, is subjected to a beat treatment step to inactivate viruses in our manufacturing process. To validate the efficacy of this inactivation, we have simulated the heat treatment procedure at a reduced scale using hemoglobin solution spiked with representative viruses. Human Immune-deficiency Virus (HIV), Cytomegalovinis (CMV), and Duck Hepatitis B Virus (DHBV) were used in this validation. Inoculation with concentrated virus was performed just prior to the heat treatment to determine the effect of that specific process step. Samples were taken before, during, and after heat treatment and assayed for virus liter in an attempt to assess the rate as well as (be extent of virus inactivation. CMV was analyzed in a plaque assay using MRC-5 indicator cells. The liter was reduced from 3.3×106 plaque forming units (PFU) per mL to < 5×101 PFU/mL (detection limit) within 30 minutes. DHBV was analyzed by inoculation of serially diluted samples into Pekin ducklings, followed at intervals by screening sera for DHBV DNA by dot blot hybridization. The liter was reduced from 5.0×104 duck infectious units (DIU) per mL to < 5×10° DIU/mL (detection limit) within 1 hour. HIV liters were determined through an ELISA assay for p24 antigen present in peripheral blood lymphocyte co-cultivation supematants. The titer was reduced from 2.0×104 infectious units (IU) per mL to < 2×10° lU/mL (detection limit) within 1 hour. These data indicate that high liters of these blood borne viruses are rapidly inactivated by this heat treatment process.

This publication has 1 reference indexed in Scilit: