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

  • Authors:
    • Adel Badran
    • Ryutaro Asano
    • Makoto Nakayama
    • Yasuhiro Watanabe
    • Takeshi Nakanishi
    • Mitsuo Umetsu
    • Hiroki Hayashi
    • Yu Katayose
    • Michiaki Unno
    • Izumi Kumagai
  • View Affiliations

  • Published online on: May 1, 2010     https://doi.org/10.3892/ijo_00000606
  • Pages: 1229-1234
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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.

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May 2010
Volume 36 Issue 5

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Badran A, Asano R, Nakayama M, Watanabe Y, Nakanishi T, Umetsu M, Hayashi H, Katayose Y, Unno M, Kumagai I, Kumagai I, et al: Target cell-restricted apoptosis induction by 528scFv-TRAIL fusion protein specific for human EGFR and expressed in Escherichia coli. Int J Oncol 36: 1229-1234, 2010
APA
Badran, A., Asano, R., Nakayama, M., Watanabe, Y., Nakanishi, T., Umetsu, M. ... Kumagai, I. (2010). Target cell-restricted apoptosis induction by 528scFv-TRAIL fusion protein specific for human EGFR and expressed in Escherichia coli. International Journal of Oncology, 36, 1229-1234. https://doi.org/10.3892/ijo_00000606
MLA
Badran, A., Asano, R., Nakayama, M., Watanabe, Y., Nakanishi, T., Umetsu, M., Hayashi, H., Katayose, Y., Unno, M., Kumagai, I."Target cell-restricted apoptosis induction by 528scFv-TRAIL fusion protein specific for human EGFR and expressed in Escherichia coli". International Journal of Oncology 36.5 (2010): 1229-1234.
Chicago
Badran, A., Asano, R., Nakayama, M., Watanabe, Y., Nakanishi, T., Umetsu, M., Hayashi, H., Katayose, Y., Unno, M., Kumagai, I."Target cell-restricted apoptosis induction by 528scFv-TRAIL fusion protein specific for human EGFR and expressed in Escherichia coli". International Journal of Oncology 36, no. 5 (2010): 1229-1234. https://doi.org/10.3892/ijo_00000606