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Cell-free production of functional antibody fragments

Cited 14 time in Web of Science Cited 16 time in Scopus
Authors
Oh, In-Seok; Lee, Ji-Chul; Lee, Myung-shin; Chung, Jun-ho; Kim, Dong-Myung
Issue Date
2010-01
Publisher
SPRINGER
Citation
BIOPROCESS AND BIOSYSTEMS ENGINEERING; Vol.33 1; 127-132
Keywords
Cell-free protein synthesis systemBotulinum neurotoxinAntigen-binding fragment (Fab)Disulfide bondAntibody
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 mu g of functional Fab was obtained after purification from 1 mL reaction mixture.
ISSN
1615-7591
Language
English
URI
http://hdl.handle.net/10371/76852
DOI
https://doi.org/10.1007/s00449-009-0372-3
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College of Medicine/School of Medicine (의과대학/대학원)Dept. of Biochemistry & Molecular Biology (생화학교실)Journal Papers (저널논문_생화학교실)
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