Effective targeted cytotoxicity of neuroblastoma cells

Patrick B. Thomas, Stephen J. Delatte, Aimee Sutphin, Arthur E. Frankel, Edward P. Tagge

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


Background/Purpose: Despite aggressive treatment with surgery, chemotherapy, and radiotherapy, the prognosis for many children with neuroblastoma remains poor. Targeted toxins represent novel cancer therapeutics designed to selectively target and kill cancer cells. The authors have developed a novel fusion toxin, DT5F11, consisting of truncated diphtheria toxin (DTA) linked to a single chain antibody (sc5F11) targeting the GD2 antigen found on most neuroblastoma cells. This report describes the construction, expression, and in vitro function of DT5F11. Methods: Utilizing restriction enzyme digestion, polymerase chain reaction amplification, and gel electrophoresis, the prkDTL5F11 plasmid was created by the fusion of distinct coding sequences for a single-chain GD2 targeting antibody (sc5F11) and truncated diphtheria toxin (DTA). DH5α Escherichi coli-competent cells were transformed with prkDTL5F11; DNA was amplified, isolated, and sequenced. The fusion protein was expressed and assayed by Western blot. Targeted cytotoxicity was analyzed on GD2-positive (SK-N-AS, IMR-32, SK-N-MC, LAN-1) and GD2-negative (HeLa) cells. Results: Fluorescent dye-labeled cycle sequencing identified the constructed fusion toxin gene. Western blot analysis using a mouse antihuman DTA antibody showed a 69-kD band identifying the fusion toxin, DT5F11. Targeted cell killing with DT5F11 was seen only in GD2 positive cells. Conclusions: This study demonstrates creation of a novel fusion toxin with effective GD2-targeted cellular toxicity. Further investigation of this fusion toxin as a therapeutic agent in the management of neuroblastoma is warranted.

Original languageEnglish (US)
Pages (from-to)539-544
Number of pages6
JournalJournal of Pediatric Surgery
Issue number3
StatePublished - 2002


  • Fusion toxin
  • GD-targeting
  • Neuroblastoma

ASJC Scopus subject areas

  • Surgery
  • Pediatrics, Perinatology, and Child Health


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