Target cell-restricted apoptosis induction by 528scFv-TRAIL fusion protein specific for human EGFR and expressed in Escherichia coli

Adel Badran, Ryutaro Asano, Makoto Nakayama, Yasuhiro Watanabe, Takeshi Nakanishi, Mitsuo Umetsu, Hiroki Hayashi, Yu Katayose, Michiaki Unno, Izumi Kumagai

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We report the preparation and functional characterization of an Escherichia coli-expressed recombinant fusion protein, 528scFv-TRAIL, specific for the human epidermal growth factor receptor (EGFR) and empowered by the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). The 528scFv-TRAIL, expressed as insoluble inclusion bodies in E. coli, was solubilized and then refolded by using a modified stepwise dialysis method. Treatment with 528scFv-TRAIL resulted in the specific binding to the cell surface of EGFR-positive cells with concomitant deployment of the TRAIL moiety to DR-5 receptor in a manner comparable to a commercially available form of recombinant TRAIL (cTRAIL). 528scFv-TRAIL, prepared by either of three refolding processes described herein, showed potent cytotoxic activity against EGFR-positive TFK-1 cell line and was superior to its parental 528scFv; a recombinant variable fragment with single specificity against human EGFR. Narrow variations in the cytotoxic potential of 528scFv-TRAIL were ascribed to manipulation of redox conditions during the refolding process. Together, our findings point to the potential value of 528scFv-TRAIL for treatment of EGFR-expressing cancers. Furthermore, preparation of 528scFv-TRAIL from insoluble aggregates in a prokaryotic cell based expression system by means of in vitro refolding introduces a feasible cost-benefit, time-efficient approach for industrial-scale production.

Original languageEnglish
Pages (from-to)1229-1234
Number of pages6
JournalInternational journal of oncology
Volume36
Issue number5
DOIs
Publication statusPublished - 2010 May

Keywords

  • Apoptosis
  • E. coli
  • EGFR
  • Fusion protein
  • TRAIL
  • scFv

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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