LARGE-SCALE CHROMATOFOCUSING-BASED METHOD FOR ISOLATING THYMOSIN BETA(4) AND THYMOSIN BETA(9) FROM BOVINE-TISSUES

A large-scale method for the isolation of thymosin beta(4) (up to 120 mg) and thymosin beta(9) (up to 40 mg) from bovine lung (up to 2 kg) w as developed. The isolation protocol included tissue homogenization in 0.4 M HClO4, centrifugation, solid-phase extraction through LiChropre p RP-18 material, chromatofocusing on polybuffer exchanger PBE 94-modi fied Sepharose and dialysis against water. The isolated products were characterized by analytical isoelectric focusing, reversed-phase HPLC, electrospray ionization mass spectrometry and amino acid analysis. Th e method developed is rapid and convenient, requires no expensive equi pment and can be used for the isolation of thymosin beta(4) and homolo gous peptides from various animal tissues.