Bioactive recombinant chicken interferon-alpha (ChIFN-alpha) was expressed in a baculovirus system. For easy purification, it was expressed as ChIFN-alpha bearing histidine hexamer (His-tag) at C-terminal, designated ChIFN-alphaHis. The expressed proteins were detected by SDS-PAGE analysis with Coomassie brilliant blue staining as around 23 and 19 kDa bands thought to be immature and matured ChIFN-alphaHis respectively. The purified ChIFN-alphaHis with a nickel chelated column showed anti-viral activity in vitro.