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Prothrombin complex concentrate (PCC) when infused into patients may produce thrombotic episodes, presumably due to the presence of some thrombogenic material. In order to prepare a safer PCC we have investigated whether the addition of purified antithrombin III (AT III) during an early stage of the isolation process would be effective in reducing thrombogenicity as assessed by the Wessler stasis model. By our method the cryosupernatant of plasma is adsorbed with DEAE-Sephadex (1.5 g/L), the unadsorbed fraction removed, the DEAE-cake washed with 0.7-1.0 plasma volume of citrate saline, followed by elution with 0.1 plasma volume of 0.03 M citrate containing 2 M NaCl and 0.25 AT III unit/ml, at pH 6.8. The eluted PCC is then concentrated and desalted by ultrafiltration, filtered, dispensed and lyophilized.  In a bench-scale experiment starting from 6 L cryosupernatant, a PCC was obtained at 43% recovery and 50-fold purification over plasma based on Factor IX activity. The recoveries of Factors II, VII and X were 44, 23 and 64% respectively. The concentrate was free of detectable thrombin and plasmin and contained about 1 unit of AT III (determined immunologically) per 25 units of Factor IX. In the Wessler stasis model this PCC was nonthrombogenic at 100 Factor IX units per kilo body weight of the rabbit. Higher doses were not tried. A control PCC prepared without the addition of AT III to the DEAE-eluting buffer was nonthrombogenic at 50, but thrombogenic at 75 Factor IX units per kilo body weight of the rabbit. Subsequent incubation with AT III did not eliminate this thrombogenicity.  These results suggest that AT III can be effective in preventing thrombogenicity in PCC but not in abolishing it if already developed.

A Simple Method For Preparation Of Nonthrombogenic Prothrombin Complex