Immobilized oxidoreductase as an additive for refolding inclusion bodies: Application to antibody fragments

Kouhei Tsumoto, Mitsuo Umetsu, Hidenari Yamada, Takahiko Ito, Satoru Misawa, Izumi Kumagai

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

Three foldases - the apical domain of GroEL (mini-chaperone) and two oxidoreductases (DsbA and DsbC) from Escherichia coli - were studied in refolding a protein with immunoglobulin fold (immunoglobulin-folded protein) that had been produced as inclusion bodies in E.coli. The foldases promoted the refolding of single-chain antibody fragments from denaturant-solubilized and reduced inclusion bodies in vitro, and also effectively functioned as alternatives for labilizing agent and oxidizing reagent in the stepwise dialysis system. Immobilization of the oxidoreductases enhanced refolding and recovery of functional single-chain antibody in the dialysis system, suggesting that immobilized oxidoreductases can be used as an effective additive for refolding immunoglobulin-folded proteins in vitro.

Original languageEnglish
Pages (from-to)535-541
Number of pages7
JournalProtein Engineering
Volume16
Issue number7
DOIs
Publication statusPublished - 2003 Jul 1

Keywords

  • Foldase
  • Immobilization
  • Immunoglobulin fold
  • Oxidoreductase
  • Refolding

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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