Production of antibody fragments in Escherichia coli.
Escherichia coli is a host widely used in the industrial production of recombinant proteins. However, the expression of heterologous proteins in E. coli often encounters the formation of inclusion bodies, which are insoluble and nonfunctional protein aggregates. For the successful production of antibody fragments, which includes single-chain variable fragments (scFvs), we describe here the modification of linker, signal, and Shine-Dalgarno (SD) sequences, the coexpression of cytoplasmic and periplasmic chaperones, and a method for fed-batch cultivation with exponential feed.
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe, Japan. katsuda@kobe-u.ac.jp
Tomohisa
T
Katsuda
Hiroyuki
H
Sonoda
Yoichi
Y
Kumada
Hideki
H
Yamaji
D000818:Animals; D061249:Batch Cell Culture Techniques / Q000379:methods*; D019149:Bioreactors / Q000382:microbiology; D002417:Cattle; D003593:Cytoplasm / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis; D018832:Molecular Chaperones / Q000378:metabolism; D019897:Periplasm / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D015203:Reproducibility of Results; D057127:Single-Chain Antibodies / Q000096:biosynthesis*; D012995:Solubility; D014170:Transformation, Genetic
D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D018832:Molecular Chaperones; D057127:Single-Chain Antibodies; D004247:DNA