Related Experiment Video
Updated: Jul 30, 2026
09:46
Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
Human pS2/trefoil factor 1: production and characterization in Pichia pastoris
R Kannan1, C Tomasetto, A Staub
1Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Centre National de la Recherche Scientifique, Université Louis Pasteur, 67404 Illkirch Cedex, C. U. de Strasbourg, France.
Protein Expression and Purification
|February 13, 2001
Summary
Recombinant human trefoil factor 1 (hTFF1) was successfully produced in Pichia pastoris yeast. This study demonstrates hTFF1
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- Human trefoil factor 1 (hTFF1), also known as hpS2, is a secreted peptide with potential roles in cell proliferation and tissue repair.
- Efficient production of recombinant hTFF1 is crucial for further functional studies and therapeutic development.
Purpose of the Study:
- To establish a robust yeast-based expression system for producing recombinant human trefoil factor 1 (hTFF1).
- To characterize the purified recombinant hTFF1 and evaluate its effect on cancer cell proliferation in vitro.
Main Methods:
- Utilized Pichia pastoris for high-level extracellular secretion of hTFF1 under the GAPDH promoter.
- Employed tangential flow filtration, ion-exchange chromatography, and preparative HPLC for protein purification.
- Confirmed protein integrity and monomeric form using Western blot analysis.
Main Results:
- Achieved large-scale secretion of intact, primarily monomeric hTFF1 in the Pichia pastoris extracellular medium.
- Demonstrated that recombinant hTFF1 significantly decreases the proliferation of HCT116 cancer cells in vitro.
Conclusions:
- Pichia pastoris is an effective host for the recombinant production of functional hTFF1.
- Recombinant hTFF1 exhibits anti-proliferative effects on HCT116 cancer cells, suggesting potential therapeutic applications.
