SOLID-PHASE SYNTHESIS OF THYMOSIN BETA(10) USING A P-CYANOTRITYL RESIN - CHEMICAL CHARACTERIZATION AND IMMUNOCHEMICAL CONTROL OF THE SYNTHETIC PEPTIDE

Thymosin beta(10) (43 amino acids) was synthesized following the Fmoc- solid-phase strategy, using a new p-cyanotrityl resin. This resin show s enhanced acid stability compared with the usual trityl ester resins, thus providing selective peptide synthesis with high yield and purity . Crude T beta(10) was purified with gel filtration and semi-preparati ve reversed-phase high-performance liquid chromatography (RP-HPLC). Th e purified synthetic peptide was shown to co-elute with recombinant T beta(10) in an analytical RP-HPLC system. The amino acid analysis data were in agreement with those reported for the literature primary stru cture of natural T beta(10). The electrospray ionization mass spectrom etry (ESIMS) data showed almost identical average mass (M(r,exp))-valu es for the synthetic or the recombinant product, which were in agreeme nt with the molecular mass calculated on the basis of the proposed pri mary structure for the peptide. Furthermore, the synthetic T beta(10) exhibited similar immunochemical characteristics compared with the rec ombinant material in an in vitro ELISA test using rabbit polyclonal an ti-T beta(10) antiserum.