S-Space College of Medicine/School of Medicine (의과대학/대학원) Dept. of Biochemistry & Molecular Biology (생화학교실) Journal Papers (저널논문_생화학교실)
Cell-free production of functional antibody fragments
- Oh, In-Seok; Lee, Ji-Chul; Lee, Myung-shin; Chung, Jun-ho; Kim, Dong-Myung
- Issue Date
- BIOPROCESS AND BIOSYSTEMS ENGINEERING; Vol.33 1; 127-132
- Cell-free protein synthesis system; Botulinum neurotoxin; Antigen-binding fragment (Fab); Disulfide bond; Antibody
- 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 mu g of functional Fab was obtained after purification from 1 mL reaction mixture.
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