title: Immobilized oxidoreductase as an additive for refolding inclusion bodies: application to antibody fragments.
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.
authors.affiliation: Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, Aoba-yama 07, Aoba-ku, Sendai 980-8579, Japan. tsumoto@mail.cc.tohoku.ac.jp
authors.forename: Kouhei
authors.initials: K
authors.lastname: Tsumoto
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Umetsu
authors.forename: Hidenari
authors.initials: H
authors.lastname: Yamada
authors.forename: Takahiko
authors.initials: T
authors.lastname: Ito
authors.forename: Satoru
authors.initials: S
authors.lastname: Misawa
authors.forename: Izumi
authors.initials: I
authors.lastname: Kumagai
mesh_terms: D018834:Chaperonin 60 / Q000378:metabolism; D003956:Dialysis; D004800:Enzymes, Immobilized / Q000378:metabolism*; D007128:Immunoglobulin Fragments / Q000378:metabolism*; D002479:Inclusion Bodies / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D020673:Protein Renaturation
chemical_list: D018834:Chaperonin 60; D004800:Enzymes, Immobilized; D007128:Immunoglobulin Fragments; D010088:Oxidoreductases