Cell-free production of functional antibody fragments

In Seok Oh, Ji Chul Lee, Myung Shin Lee, Jun Ho Chung, Dong Myung Kim

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

Botulinum neurotoxin serotype B (BoNT/B)-specific Fab was expressed in a cell-free protein synthesis system derived from an E. coli extract. The cell-free synthesized antibody fragment was found to be effective in neutralizing the toxicity of BoNT/B in animal studies. Expression of functional Fab required an appropriately controlled and stably maintained redox potential. Under an optimized redox condition, the cell extract, whose disulfide reducing activity had been exhausted, could generate bio-functional Fab molecules. Use of a cell extract enriched with molecular chaperones (GroEL/ES) and disulfide bond isomerases were effective in obtaining larger quantities of functional Fab. Under the optimized reaction conditions, approximately 30 μg of functional Fab was obtained after purification from 1 mL reaction mixture.

Original languageEnglish
Pages (from-to)127-132
Number of pages6
JournalBioprocess and Biosystems Engineering
Volume33
Issue number1
DOIs
StatePublished - 1 Jan 2010

Keywords

  • Antibody
  • Antigen-binding fragment (Fab)
  • Botulinum neurotoxin
  • Cell-free protein synthesis system
  • Disulfide bond

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