Immobilised metal affinity chromatography (IMAC) beads for lysozyme separation: Synthesis and characterization study [Kromatografi afiniti logam dipegun (IMAC) untuk pemisahan lysozim: Sintesis dan kajian pencirian]

Abstract

Immobilized metal affinity chromatography (IMAC) has been established as a highly specific chromatographic technique for the production of enzymes and proteins including lysozyme. This study aimed to prepare and characterize the IMAC beads for lysozyme separation. Silica gel served as chromatographic matrix which has been coated with chitosan layer and crosslinked with glutaraldehyde (GTA-CTS-SiO2) since it is very convenient to promote fixation. Various IMAC ligands were immobilized by chelating 1500 mg/l Cu2+, Zn2+, Fe2+, Fe3+ and Al3+ ions, respectively on GTA-CTS-SiO2. IMAC which was immobilized with 1200mg/l Cu2+ exhibited the maximal immobilization capacity (27.08mg/g) of within 30 minutes incubation time. This fundamental study can be a momentous pathway to develop an efficient chromatographic system for lysozyme separation in the future