Direct recovery of clotting factor IX from unclarified human plasma by expanded bed ion exchange chromatography

In this study, STREAMLINE DEAE adsorbent was used for the direct recovery of clotting factor IX (FIX) from unclarified human plasma in expanded bed adsorption (EBA). The equilibrium adsorption studies for general protein showed that the experimental data followed the Langmuir isotherm model. The maximum adsorption capacity for general protein was 35.33 mg/ml with a dissociation constant of 0.18 mg/ml. However, the adsorption of FIX followed the Henry's law with Henry's constant of 59.78. Optimal condition for the elution of FIX with step changes of increasing concentration of NaCl was determined in small packed bed experiments conducted with clarified plasma. The results showed 94.28% of the bound FIX could be eluted with 0.7 M NaCl. The influence of liquid velocities of 100-300 cm/h on the selective adsorption of FIX from clarified plasma was also investigated in packed bed experiments. The results showed that the rate of FIX breakthrough was found to be very similar. In this work, operation at a constant liquid velocity of 200 cm/h in an expanded bed recovery process was chosen for the purification of FIX. The FIX was directly recovered using 0.7 M NaCl from unclarified plasma with a purification factor of 14 and a yield of 90% in a single step.