Increased refolding yield of disulfide bond bridged fab-toxin homodimers by the insertion of CH3 domains

Jeongwha Song, Jaeseon Won, Yongchan Lee, Muhyeon Choe

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Recombinant antibody-toxin is a bifunctional protein that binds and kills a target cell expressing a specific antigen on the surface of the cell, and its structure is chimeric, in which a toxin is fused to an antigen-binding domain such as scFv or Fab. Divalent antibody-toxin molecules showed higher cytotoxicities against cancer cell lines than monovalent molecules. However, the yields of the divalent molecules were very low. In this study, we introduced the CH2, CH3, or CH2-CH3 (=Fc) domain of antibody in the middle of the Fab-toxin between the hinge region of human IgG1 and the toxin domain to increase the yield. The covalently bonded dimer could be formed by three disulfide bridges from cysteine residues in the hinge region. The molecule with the CH3 domain showed about 3-fold higher dimerization yield than previously constructed Fab-toxin molecules, while maintaining the cytotoxic activity comparable to that of scFv-toxin. However, the introduction of CH2 or Fc domain to the same position showed little effect on the dimerization yield. We also observed that the introduction of the CH3 region made it possible to form noncovalently associated dimer molecules.

Original languageEnglish
Pages (from-to)1104-1110
Number of pages7
JournalJournal of microbiology and biotechnology
Volume16
Issue number7
Publication statusPublished - 2006 Jul

Keywords

  • Antibody refolding
  • B3 antibody
  • Constant domain of heavy chain
  • Cytotoxicity
  • Dimer
  • Divalent antibody-toxin
  • Pseudomonas exotoxin A.
  • Recombinant antibody-toxin

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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