Purification of human epidermal growth factor (urogastrone) from urine

Human epidermal growth factor has been isolated from a concentrated chromatographic eluate of human urine. The purification method utilizes six chromatographic steps including adsorption to aminoethylcellulose (AE-11), gel filtration on Sephadex G-50, carboxymethylcellulose (CM-52) chromatography, ion-exchange HPLC and reverse-phase HPLC. The final product appears homogeneous and identical to pure gamma-urogastrone when analyzed by polyacrylamide gel electrophoresis and reverse-phase HPLC using two eluent systems. The yield of the method described above allowed the development of a sensitive radioimmunoassay system for this growth factor.