Two-step chromatographic procedure for the preparation of hen egg white ovotransferrin

A two-step chromatographic procedure for the preparation of hen egg white ovotransferrin by using an anion exchanger is described. The method consists of a preliminary step of frontal chromatography, also called displacement chromatography, on a Q Sepharose Fast Flow column. This simple and powerful chromatographic step enables the pre-purification of ovotransferrin that is further purified by a second conventional preparative chromatography performed on the same Q Sepharose FF column. The ovotransferrin obtained is found to be homogeneous in electrophoresis and analytical chromatography. Its purity rate is calculated at 97.5% and its amino acid composition is close to that obtained from the nucleotide sequence of ovotransferrin. Moreover, ovotransferrin is iron-free, thus suitable for studies in relation to its metal-binding activity.