Human pS2/trefoil factor 1: Production and characterization in Pichia pastoris

The recombinant protein human trefoil factor 1 (hTFF1), formerly called hpS 2, has been produced for the first time in a yeast-based expression in Pich ia pastoris. hTFF1 was secreted in large amounts in the extracellular mediu m of P, pastoris under the control of the glyceraldehyde-3-phosphate dehydr ogenase promoter. The fermentation broth containing hTFF1 was concentrated by tangential flow filtration prior to purification by anion- and cation-ex change chromatography, followed by preparative high-performance liquid chro matography. The resulting hTFF1 was found to be intact by Western blot anal ysis. Further analysis revealed mainly the presence of the monomeric form o f the hTFF1 peptide. Finally, in vitro, the recombinant hTFF1 was shown to decrease proliferation of the HCT116 cancer cells, (C) 2001 Academic Press.