Comparison of affinity membrane adsorbers for the purification of recombinant human erythropoietin (rhEPO)

In this work affinity membrane adsorbers were investigated for the chromatographic purification of recombinant human erythropoietin (rhEPO) produced in mammalian cells. Cibacron Blue (CB), IDA‐Cu+2, wheat germ agglutinin (WGA), concanavalin A (ConA) and an anti‐EPO monoclonal antibody (MAb) were tested as affinity ligands, attached to microporous Sartobind® membranes. In experiments carried out with cell culture supernatant, the best results were obtained with Sartobind–CB, Sartobind–WGA and Sartobind–MAb membranes. The thermodynamic parameters were determined by adsorption isotherms of rhEPO onto the membranes. Sartobind–ConA presented the lowest affinity for rhEPO, as evidenced by a lower association constant. For Sartobind–CB, Sartobind–IDA‐Cu+2 and Sartobind–MAb KA was in the order of 105 L mol−1, whereas for Sartobind–WGA it was 106 L mol−1. Sartobind–CB eluates were also investigated by RP‐HPLC. The purity level achieved in this one‐step purification strategy was 55%, indicating that the Sartobind–CB membrane is a promising affinity membrane for rhEPO purification.