Engineering of human Fas ligand extracellular domain (hFasLECD) protein can provide us with an advanced way for treatment or diagnosis of many serious diseases represented by cancers. In this study, we prepared a site-specific biotin group conjugate of hFasLECD and conducted the characterization of its cell-death-inducing activity. Precipitation of complexes using magnetic beads for affinity-based capturing revealed that the conjugate retained the original binding activity to both human Fas receptor extracellular domain and streptavidin. A significant sensitization effect on the cell-death-inducing activity against a colorectal cancer cell line, HT-29 cells, was observed for the pretreatment with human interferon-γ, accompanied by a synergistic effect of 5-fluorouracil. The findings suggested that the prepared hFasLECD conjugate would be applicable to the future development of medically useful devices to detect the counterpart receptors in biological specimens as well as novel cytotoxic agents against diseased cells.
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