Recombinant expression and downstream processing of the disulfide‑rich tumor‑targeting peptide chlorotoxin

  • Authors:
    • Xiao‑Min Wang
    • Xiao Luo
    • Zhan‑Yun Guo
  • View Affiliations

  • Published online on: July 24, 2013     https://doi.org/10.3892/etm.2013.1234
  • Pages: 1049-1053
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Abstract

Chlorotoxin (CTX) is a scorpion‑derived disulfide‑rich peptide that targets malignant tumors by binding the cell surface matrix metalloproteinase‑2 and annexin A2. Various CTXs labeled with functional moieties have shown great potential for tumor diagnosis and treatment. In the present study, we established an efficient approach for preparing mature CTX that may be used for experimental and therapeutic purposes. The designed CTX precursors carried either a 6xHis‑tag or a 6xHis‑tag and a glutathione transferase (GST)‑tag and were recombinantly expressed in Escherichia coli. Following S‑sulfonation, the precursors were purified using immobilized metal‑ion affinity chromatography. Subsequent to the removal of the tag by enterokinase cleavage and in vitro oxidative refolding, mature CTX was obtained with a considerable yield. The yield of mature CTX whose precursors carried a 6xHis‑tag and a GST‑tag (2 mg per liter of culture) was ~10‑fold that of the mature CTX whose precursors carried a 6xHis‑tag (150‑200 µg per liter of culture). The folded CTX inhibited the migration of glioma cells in a concentration‑dependent manner, suggesting it was biologically active.
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October 2013
Volume 6 Issue 4

Print ISSN: 1792-0981
Online ISSN:1792-1015

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Spandidos Publications style
Wang XM, Luo X and Guo ZY: Recombinant expression and downstream processing of the disulfide‑rich tumor‑targeting peptide chlorotoxin. Exp Ther Med 6: 1049-1053, 2013
APA
Wang, X., Luo, X., & Guo, Z. (2013). Recombinant expression and downstream processing of the disulfide‑rich tumor‑targeting peptide chlorotoxin. Experimental and Therapeutic Medicine, 6, 1049-1053. https://doi.org/10.3892/etm.2013.1234
MLA
Wang, X., Luo, X., Guo, Z."Recombinant expression and downstream processing of the disulfide‑rich tumor‑targeting peptide chlorotoxin". Experimental and Therapeutic Medicine 6.4 (2013): 1049-1053.
Chicago
Wang, X., Luo, X., Guo, Z."Recombinant expression and downstream processing of the disulfide‑rich tumor‑targeting peptide chlorotoxin". Experimental and Therapeutic Medicine 6, no. 4 (2013): 1049-1053. https://doi.org/10.3892/etm.2013.1234