Cell-free production of functional antibody fragments

被引：0
作者：
In-Seok Oh
Ji-Chul Lee
Myung-shin Lee
Jun-ho Chung
Dong-Myung Kim
机构：
[1] Seoul National University,Institute of Molecular Biology and Genetics
[2] Abxign,Department of Microbiology and Immunology
[3] Inc.,Department of Biochemistry and Molecular Biology
[4] Eulji University School of Medicine,Department of Fine Chemical Engineering and Applied Chemistry
[5] Cancer Research Institute,undefined
[6] Seoul National University College of Medicine,undefined
[7] Chungnam National University,undefined
来源：
Bioprocess and Biosystems Engineering | 2010年 / 33卷
关键词：
Cell-free protein synthesis system; Disulfide bond; Antibody; Antigen-binding fragment (Fab); Botulinum neurotoxin;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
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.
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页码：127 / 132
页数：5
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