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

Wang,Xiao‑Min; Luo,Xiao; Guo,Zhan‑Yun

doi:10.3892/etm.2013.1234

October 2013 Volume 6 Issue 4

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October 2013 Volume 6 Issue 4

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Article

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

Authors:
- Xiao‑Min Wang
- Xiao Luo
- Zhan‑Yun Guo
View Affiliations / Copyright

Affiliations: Institute of Protein Research, College of Life Sciences and Technology, Tongji University, Shanghai 200092, P.R. China
Pages: 1049-1053
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Published online on: July 24, 2013

https://doi.org/10.3892/etm.2013.1234
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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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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

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

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