Stable supply of large amounts of human Fab from the inclusion bodies in E. coli

Testuro Fujii, Takatoshi Ohkuri, Reiko Onodera, Tadashi Ueda

Research output: Contribution to journalArticle

16 Citations (Scopus)


Recombinant human Fab-H chain and L chain were separately expressed as inclusion body using Escherichia coli. After solubilization of Fab-H chain and L chain by the reduction and S-alkyldisulphidation in 8 M urea, about 100 mg of purified Fab-H chain and about 160 mg of L chain could be obtained from 11 of each culture by ion-exchange chromatogram in the presence of 8 M urea. Combination of the lyophilized Fab-H chain and L chain could be efficiently folded to native human Fab by using the stepwise dialysis method and the human Fab was purified with cation-exchange chromatogram. In the folding procedure, it was found that cysteamine and cystamine with positive charge were effective to improve the folding yield of human Fab. Moreover, from comparison of folding yield in the presence of ten kinds of additives, it was suggested that taurine was effective to improve the folding of human Fab. Consequently, we could obtain about 60 mg of folded human Fab from 11 of each culture under the optimum conditions.

Original languageEnglish
Pages (from-to)699-707
Number of pages9
JournalJournal of Biochemistry
Issue number5
Publication statusPublished - May 1 2007


All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this