By using ion-exchange preparative chromatography (IEPC) and reversed-phase high performance liquid preparative chromatography (RP-HPLPC), thymosin α1 was isolated and purified from the crude product synthesized by the solid-phase peptide synthesis (SPPS) method. The purity of the final product reached 95% through ion-exchange chromatography on DEAE Sepharose Fast Flow chromatography and Delta-Pak^TM C18 purification after optimizing the chromatographic conditions. The capacity of purification process was 50mg/circle. The total yield was 36%. The technology is simple and reliable, and can be scaled up easily.