Novel immobilized Cu+2 ion grafted cellophane membranes for affinity separation of His-Tag Chitinase

Abstract

Two kinds of immobilized Cu+2 ion grafted cellophane membrane were prepared using methacrylic acid (MAA) and methyl methacrylate (MMA). The affinity grafted membranes were tested in separation of His-Tag Chitinase enzyme from BSA-protein mixture. PMAA grafted membranes show a higher affinity than PMMA grafted ones towards the separation of Chitinase enzyme. Both types of affinity membranes show almost the same desorption capability for both proteins with small advantage for PMAA grafted affinity membranes. No leakage of Cu+2 ions was detected during the protein elution process. The characters of the obtained Cu+2 immobilized grafted membranes recommended them as novel immobilized metal affinity membranes (IMAMs) for His-Tag protein separation and presenting a solution for the metal ion leakage problem; the main reason for limiting the use of metal ion affinity chromatography technique.