Enhanced formation of disulfide-bridged dimer (Fab-PE38)2 utilizing repeats of the Fab binding domain of protein G

YongChan Lee, JongChan Park, SoonWook Kweon, MuHyeon Choe

Research output: Contribution to journalArticle

Abstract

Fab-PE38 used in this study is B3(Fab)-ext-PE38, and it is an antibody toxin that is made by fusing the Pseudomonas exotoxin to the Fab domain of B3 antibody. This antibody toxin selectively binds to cancer cells and kills the target cancer cells. B3(Fab)-ext-PE38 has a cysteine residue on the ext sequence, and (B3(Fab)-ext-PE38)2 is the disulfide-bridged dimer of the B3(Fab)-ext-PE38 monomer. (B3(Fab)-ext-PE38)2 has been found to have 11-fold higher cytotoxicity on the CRL-1739 cell line than monomeric B3(scFv)-PE38. We made a recombinant tandem repeat of the domain III of Streptococcal protein G that has Fab binding property up to seven repeats. Multiple monomers were found to form non-covalent complexes with this tandem repeat. Complexes were purified by size-exclusion chromatography, and we could enhance the production of the disulfide-bridged dimer by reduction and oxidation of the complexes. The tandem repeat makes close intermolecular interactions between monomers possible, and the use of it greatly enhances the yield of the disulfide-bridged dimer.

Original languageEnglish
Pages (from-to)5127-5131
Number of pages5
JournalJournal of Biological Chemistry
Volume285
Issue number8
DOIs
Publication statusPublished - 2010 Feb 19

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Tandem Repeat Sequences
Disulfides
Dimers
Carrier Proteins
Monomers
Cells
Antibodies
Exotoxins
Proteins
Size exclusion chromatography
Cytotoxicity
Pseudomonas
Gel Chromatography
Cysteine
Neoplasms
Cell Line
Oxidation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Enhanced formation of disulfide-bridged dimer (Fab-PE38)2 utilizing repeats of the Fab binding domain of protein G. / Lee, YongChan; Park, JongChan; Kweon, SoonWook; Choe, MuHyeon.

In: Journal of Biological Chemistry, Vol. 285, No. 8, 19.02.2010, p. 5127-5131.

Research output: Contribution to journalArticle

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