Large‐Scale Preparation of a Highly Purified Solvent‐Detergent Treated Factor VIII Concentrate

Abstract

Large-scale adaptation of a recently reported glycine precipitation method for the production of factor VIII (FVIII) concentrate is described. Scaling up of the method required some modification including the addition of aluminum hydroxide to the glycine buffer to reduce the level of contaminating proteins in the final preparation and the use of centrifugation to replace filtration by glass beads. Furthermore, the resultant product was virus inactivated by incorporation of the organic solvent and detergent technique. At industrial level, the modified method gave a good recovery of FVIII activity (230 IU/l plasma) with high purity (4 IU/mg protein). The final product, after virus inactivation and lyophilization, yielded 185 IU of FVIII activity per liter of starting plasma and was considered to be suitable for clinical evaluation.

Progress in large‐scale purification of factor VIII/von Willebrand factor concentrates using ion‐exchange chromatography

“…Early attempts to purify FVIII from human plasma for therapeutic use were largely unsuccessful [4]. First products were obtained through precipitation with different agents but had a very low purity [5,6]. The introduction of chromatographic purification steps in the manufacturing processes allowed to obtain high‐purity concentrates.…”

Effect of Monosaccharides during Severe Dry Heat Treatment of Coagulation Factor VIII Concentrates

“…For pasteurization, high concentrations of stabilizers are needed during heating in the liquid state in order to ensure survival of the labile factor VIII protein, and these have to be removed from the product after-wards [4]. The application of the solvent detergent method needs a particle-free solution, and subsequent removal of the chemicals used for the inactivation of lipid enveloped viruses [5].…”

“…wards [4], The application of the solvent detergent meth od needs a particle-free solution, and subsequent removal of the chemicals used for the inactivation of lipid enve loped viruses [5], Severe dry heat treatment (80°C for 72 h) has a good safety record with regard to the lack of transmission of non-A non-B hepatitis and the hepatitis C virus in coag ulation factor concentrates [6,7], It is the only method that can be applied to the product in the final container [8,9], Most processing steps inflluence the effect of this severe thermal treatment on the characteristics of the final product ( fig. 1).…”

Preparation of Intermediate- Purity Factor VIII Concentrate by Direct Gel Filtration of Cryoprecipitate

“…Each method gave a different recovery of FVIII in their cryoprecipitate from plasma, hence the table only compares the increased purification relative to cryoprecipitate. Benny et al [9] used the CPG method of Margolis and Rhoades [2] whereas the rest [7,15,16] were selective precipitation methods. In Dengler et al [7], the large increase in puri fication factor is primarily due to the use of S-400HR in the last desalting step.…”