A Divalent Recombinant Immunotoxin Formed by a Disulfide Bond between the Extension Peptide Chains

Ji Hae Park, Hye Won Kwon, Hee Kyoung Chung, Ik Hwan Kim, Kwangseog Ahn, Eui Ju Choi, Ira Pastan, Mu Hyeon Choe

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)


Recombinant immunotoxin for the treatment of cancer was made by connecting toxins to 'carcinoma-specific' antibodies that selectively bind to cancer cells, then kills them without harming the normal cells. The divalent recombinant immunotoxin, [B3(Fab)-ext-PE38]2, is a derivative of B3(Fab)-PE38. B3(Fab)-PE38 was made by fusing the Fab domain of the monoclonal antibody (MAb) B3 to PE38, a truncated mutant form of Pseudomonas exotoxin (PE). In this study, B3(Fab)-ext-PE38 was constructed, which has the hinge region of the B3(Fab)-PE38 extended with the peptide extension, G4C(G4S)2, and connected to the C3 connector. The Cys residue of the extension peptide chain makes the disulfide bond between the two Fab domains. The extension sequence (ext) makes the dimerization of B3(Fab)-ext-PE38 easier to form the divalent immunotoxin, because it decreases the steric hindrance between the two PE38s. The constructed genes were expressed in E. coli as inclusion bodies. Polypeptides that were obtained from the inclusion body were refolded, and the active forms were purified. The ID50 values of the divalent molecule, [B3(Fab)-ext-PE38]2, were about 4 ng/ml on A431 cell lines, about 1 ng/ml on CRL1739 cell lines, and 5 ng/ml on MCF-7 cell lines. The [B3(Fab)-ext-PE38]2 showed about a 12-fold higher cytotoxicity on CRL1739 cell lines than B3(scFv)-PE40 did.

Original languageEnglish
Pages (from-to)398-402
Number of pages5
JournalMolecules and cells
Issue number3
Publication statusPublished - 2001 Dec 31


  • Antibody Refolding
  • B3 Antibody
  • Cytotoxicity
  • Divalent Immunotoxin
  • Pseudomonas Exotoxin A
  • [B3(Fab)-ext-PE38]

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology


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