Document Detail

Cell-free production of functional antibody fragments.
MedLine Citation:
PMID:  19701777     Owner:  NLM     Status:  MEDLINE    
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 microg of functional Fab was obtained after purification from 1 mL reaction mixture.
In-Seok Oh; Ji-Chul Lee; Myung-shin Lee; Jun-ho Chung; Dong-Myung Kim
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-08-23
Journal Detail:
Title:  Bioprocess and biosystems engineering     Volume:  33     ISSN:  1615-7605     ISO Abbreviation:  Bioprocess Biosyst Eng     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2009-12-21     Completed Date:  2010-03-02     Revised Date:  2013-11-25    
Medline Journal Info:
Nlm Unique ID:  101088505     Medline TA:  Bioprocess Biosyst Eng     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  127-32     Citation Subset:  IM    
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MeSH Terms
Antibodies, Monoclonal / biosynthesis*,  genetics,  immunology
Botulinum Toxins / immunology
Cell-Free System
Escherichia coli / metabolism
Immunoglobulin Fab Fragments / biosynthesis*,  genetics,  immunology
Recombinant Proteins / biosynthesis*,  genetics,  immunology
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Immunoglobulin Fab Fragments; 0/Recombinant Proteins; 0Y70779M1F/rimabotulinumtoxinB; EC Toxins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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