As by-products of chicken slaughtering, chicken feathers are produced and mostly discarded without proper treatment, which results in serious environment pollution. Therefore, the appropriate treatment and utilization of chicken feathers are needed. In particular, chicken feathers can be used as protein sources for the preparation of protein hydrolysates, considering that chicken feathers have a large amount of proteins. In this study, chicken feather protein hydrolysates were prepared and their iron-binding peptides were isolated. Chicken feather protein was extracted from feathers of slaughtered chicken, and its hydrolysates were prepared via hydrolysis with Flavourzyme for 8 h. Then the chicken feather protein hydrolysates were ultra-filtered to obtain small peptide fractions and fractionated using Q-Sepharose and Sephadex G-15 columns to isolate their iron-binding peptides. Two major fractions were produced from each of the Q-Sepharose ion exchange chromatography and the Sephadex G-15 gel filtration chromatography. Among the fractions, the peptide fraction with a high iron-binding activity level, F12, was isolated. In addition, isolation of iron-chelating peptide purified from spirulina protein hydrolysates was investigated. Spirulina protein was obtained by ammonium sulfate precipitation, and hydrolyzed using Alcalase and Flavourzyme. The degree of hydrolysis was determined by trinitrobenzene sulfonic acid (TNBS) assay and SDS-PAGE, and spirulina protein hydrolysates were ultra-filtered below 3 kDa. The ultra-filtered hydrolysates were fractionated by Q-Sepharose fast flow and Sephadex G-15 column, and the iron-chelating activity of each fraction was determined. The fraction with the highest iron-chelating activity was identified by MALDI-TOF/TOF mass spectrometry (Matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry). The amino acid sequence of the iron-chelating peptide was identified to be Thr-Asp-Pro-Ile(Leu)-Ala-Ala-Cys-Ile(Leu) which has a molecular weight of 802.423 Da. In conclusion, the iron-chelating peptide could be used as an iron supplemental material in functional industry.