Development and Small-Scale Production of a Severely Heated Factor VIII Concentrate

Abstract

The small-scale production of a severely heated factor VIII concentrate is described. As starting material for the production 48 units of fresh frozen plasma from whole-blood donations or 24 units of plasma from apheresis are used. During a glycine and sodium chloride fractionation, contaminating proteins are removed from a pooled extract of single-donor cryoprecipitates. The resulting factor VIII-rich precipitate is redissolved in freeze-drying buffer and this solution is desalted by diafiltration. After filtration, this solution is dispensed into 10 vials and frozen. The product is freeze dried, and heat treated at 80°C for 72 h. The mean yield of the heat-treated product improved from 160 IU factor VIII per liter plasma at the start of the production to 215 IU per liter plasma after a modification of the purification process. The validation of the virus inactivation during freeze-drying and 80°C heat treatment for 72 h showed a reduction of ≥ 7.6 log PFU/ml for Sindbis and a reduction of ≥ 6.4 log TCID(50)/ml for HIV-1. The factor VIII concentrate was clinically tested in 6 patients. It possessed a normal half-life, showed a high recovery and no side effects. A Dutch license was granted in June 1991 and since then this product has been routinely manufactured in three Dutch blood banks.

Allometry of Factor VIII and Informed Scaling of Next-Generation Therapeutic Proteins

“…The complete set of collected data is summarized in Table 1. A total of 60 publications were included representing mice, dogs, and humans with HA as well as normal mice, rats, rabbits, and monkeys 7–11,14–16,25–76. A summary of the NCA derived PK parameters CL, V ss , MRT, and half‐life ( t 1/2 ) are presented in Table 2.…”

Preparation and viral inactivation of cryoprecipitate in blood banks in resource‐limited countries

“…The resulting FVIII-rich precipitate is re-dissolved in a formulation buffer, diafiltrated, dispensed in vials and freeze-dried. The product is then heat-treated at 80 ° C for 72 h. Such heat treatment was shown to inactivate ≥ 6·4 log and ≥ 7·6 of HIV and Sindbis virus, respectively [26]. Petersen and Bird [27] have provided operational details of such a closed bag system procedure that further requires transfer into bottles for freeze-drying.…”

Concentrate safety and efficacy

“…If heat is applied while the concentrate is still in solution (pasteurization), then the temperature (60 °C) and time (10 h) needed to kill both HIV and hepatitis viruses are less than if steam‐vapour heat is used after lyophilization (a technique applied only at the former Immuno plant in Vienna) or if the final vial of lyophilized concentrate is baked (dry heat). In the latter instance, temperatures of 80 °C for 72 h [5] or 100 °C for 30 min [6] have been successful in killing hepatitis viruses.…”