Thymosin α 1 was synthesized manually. The washing condition and the test of the degree of coupling were studied. In addition, the analytical method of crude product was set up. Thymosin α 1 was synthesized with the solid-phase method, by using the base-liable Fmoc group to protect the α-amino acid and N,N′-diisopropylcarbodiimide (DIC) as the coupling regent. The crude product was analyzed by reversed-phase high performance liquid chromatography, SDS-PAGE and liquid chromatography/mass spectroscopy.The synthesis yield was 84.89%. The analytical results indicated that the crude product contained much thymosin α 1, and the purity was 50.3%. DIC solid-phase chemical synthesis of thymosin α 1 was applicable, and the yield was improved greatly compared to other known methods.
By using ion-exchange preparative chromatography (IEPC) and reversed-phase high performance liquid preparative chromatography(RP-HPLPC), thymosin α 1 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.
