Affinity adsorption of human vitamin K-dependent coagulation factor IX onto heparin-like poly (styrene sodium sulfonate) adsorbent

Adsorption of proteins on polymer material plays an important role in a number of fields, particularly in separation of biomolecules by chromatographic methods. The work reports here the synthesis of modified cross-linked polystyrene gel beads as a stationary phase in liquid chromatography for the purification of factor IX. Suitable chemical groups, such as sulfonate which confer this polymer heparin-like adsorbing property, were grafted on the aromatic ring of the hydrophobic matrix. This functional group was chosen on the basis of the biospecific molecular interactions between factor IX and its ligand particularly heparin in such manner to enhance its binding ability and efficacy. Adsorption of factor IX on to this functional polymer was performed under physiological conditions according two modes: non-competitive adsorption (adsorption of factor IX alone) and competitive adsorption (adsorption of factor IX in the presence of another vitamin-K dependent coagulation factors). The adsorbed factor IX content at the interface allows to establish the chemisorption isotherm curves. The adsorption rate in both cases was found to be significantly high and the affinity constants, estimated by the Langmuir model, were: 4.7 × 10⁸ and 4.1 × 10⁸ l/M respectively. Affinity chromatography on column using this functional polymer as a stationary phase confirms its high ability to adsorb factor IX at low ionic strength. Thus, the synthesized packing material gel functionalised by sulfonate group can be used advantageously as a heparin-like adsorbent in purification of factor IX.