Pemurnian Rekombinan Eritropoietin Manusia (rhEPO) Menggunakan Kromatografi Afinitas Ni-NTA dan Co-Talon

Abstract

Suitable and efficient techniques for purification of recombinant human erythropoietin (rhEPO) are required to obtain pure rhEPO which can be continued to preclinical trials step. One of these techniques is immobilized metal affinity chromatography (IMAC), which has advantages such as high selectivity, easy to elute, and simple procedure for column regeneration. At least, there are 2 kinds of commercial IMAC resin using nitriloacetic acid (NTA) matrix as chelating agent, namely nickel-based (Ni-NTA) and cobalt-based IMAC resin (Co-NTA), the latter is well known as Talon. This research studied the influence of metal ions Ni2+ and Co2+ in rhEPO purification. The results indicated that Ni- NTA had higher affinity in binding rhEPO protein compared with Co-Talon. Average total protein by Ni-NTA was 226.72 μg/mL and Co-Talon was 60.52 μg/mL, but the protein purity with Co-Talon was better than Ni-NTA. Base on protein yield that can be separated, Ni-NTA is more suitable to obtain high yield of rhEPO protein.