An immunoaffinity chromatography (IAC) column was prepared and optimized to purify melamine in animal-derived foods followed by determination with a liquid chromatography method. The column showed high binding capacity of 1,250 ng for melamine with recovery higher than 96%, and the column could be renewed and repeatedly used for at least 30 cycles. The extracts of chicken, egg and milk samples were directly loaded onto IAC column for purification, and the analyte retained on the column was eluted with 4 mL of modified Tris buffer without carryover. The purification effect was satisfactory and better than that of the conventional solid phase extraction. Recoveries from the melamine-fortified blank samples were between 82.5 and 101.4% with coefficient of variation (CV) lower than 10.6%.