Immobilized metal-chelated affinity chromatography(IMAC) membranes were prepared for separating a recombinant fusion antibacterial peptide,which carried a polyhistidine sequence (HIS₆-tag) at the N-terminus. Low bleeding of metal ion was achieved. It was proved that the properties of IMAC membranes were better than conventional chelating sepharose fast flow column. The fractionation of recombinant proteins that carry a polyhistidine tag is currently perhaps the most promising application of IMAC.