Isolation and identification of thymosin α-1 from calf spleen using high performance liquid chromatography

A peptide isolated from calf spleen has been identified as thymosin α1. The isolation involved defatting of the desiccated glands with acetone, extraction of the acetone power with pyridine acetate pH 5.5, heat denaturation, reverse phase chromatography on an RP-8 column, anion exchange chromatography on a Partisil SAX column and, finally, reverse phase purification on a μBondapak C18 column. The identification was based on: 1) amino acid analysis; 2) thermolysin digest; and 3) retention time in two different HPLC systems. The amount isolated from the spleen was 10–20% of that isolated from calf thymus glands.