Studies on the Stability of VIII:C during the Manufacture of a Factor VIII Concentrate for Clinical Use¹

Abstract

The stability of VIII:C was investigated by monitoring samples taken at different points from a routine process for the manufacture of factor VIII concentrate and by examining the stabilising influence of a number of product formulations. Loss of VIII:C over process-finishing procedures (formulation, 0.22 micron filtration, dispensing) was associated with a citrate-induced inactivation which could be prevented by controlling the ionised calcium concentration of the solution. These results were obtained using a one-stage clotting assay but were not observed using a two-stage assay. No evidence for activation was found in vitro (e.g. by FPA generation and VIII:C stability) and the yield increase suggested by the one-stage assay was supported by results from a controlled clinical evaluation.

Large‐Scale Gel Filtration Chromatography for the Production of a Solvent/Detergent‐Treated High‐Purity Factor VIII Concentrate

“…Initial values for soherddetergent treatment are 2 g/l (TNBP) and er for further reduction of ionic interactions did not have any beneficial effect on FVIII : c recovery. Adding 0.002 mol/l Ca2+ to the elution buffer seemed to improve the yield (about 5%); however, this may only be an artifact of the one-stage clotting assay [19].…”

The manufacture of blood plasma products in Scotland: a brief history

“…Heat treatment of Factor VIII concentrate began in November 1984, within days of a report that HIV might be inactivated by this procedure and with stocks obtained from donations obtained from late-1983 being heat treated. Calcium was added to stabilise factor VIII during processing, 11 a technique now widely used in the preparation of both recombinant and plasma-derived Factor VIII concentrates. Further research led to the introduction of a new Factor VIII concentrate in 1987 which could be heated at an even higher temperature, 12 and eventually confirmed as effective against hepatitis C virus (HCV).…”

Direct Capture of Plasma Factor VIII:C by Ion Exchange Chromatography

“…Citrate may help to stabilise FVIITC although this may not be the sole factor involved since a similar poor result as when using PBS buffer was achieved using a 20 mM Tris, 10 mM citrate, saline buffer (data not shown). The presence of 1 mM CaCl2 may be the factor in determining FVIITC stability [24,25].…”