EFFECT OF MONOSACCHARIDES DURING SEVERE DRY HEAT-TREATMENT OF COAGULATION-FACTOR-VIII CONCENTRATES

During product development of a factor VIII concentrate (Dutch blood b anks) the conversion from unsterilized to autoclaved freeze-drying buf fer caused impaired product characteristics after severe dry heat trea tment (80 degrees C for 72 h). Analysis of the freeze-drying buffers s howed the presence of fructose and glucose in heated buffers, resultin g from hydrolysis of sucrose. The detrimental effect of glucose and fr uctose on solubility, yield of factor VIII and color of the heat-treat ed product was confirmed by freeze-drying and heating products spiked with increasing levels of these monosaccharides. The effect of the use of freeze-drying buffers autoclaved with and without sucrose was exam ined in two other factor VIII concentrates, S(8) and Z8 (Protein Fract ionation Centre, Edinburgh, UK). If sucrose was present during autocla ving of the buffer, a slightly lower yield over freeze-drying and 80 d egrees C heat treatment was observed. Since glucose is present as a su bstrate in the medium for the host cells during cultivation of viruses , its potential effect on the 80 degrees C heated product (Dutch blood banks) was examined during the validation study of the inactivation o f HIV-1 and Sindbis. The cultivation cycles for the virus inolcula wer e simulated and residual glucose levels measured. In the supernatant m edium of the host cell culture used for the propagation of Sindbis no residual glucose was found. Glucose however was found in the supernata nt medium of the host cell culture for propagation of HIV-1, and there fore small molecular weight substances were removed from the actual HI V-1 inoculum by ultrafiltration. The study of the inactivation of both viruses during freeze-drying and 80 degrees C heat treatment was subs equently carried out without complications.