Co-operative effects of protein engineering and vector optimization on high yield expression of functional bivalent miniantibodies in Escherichia coli.
The volumetric yield of functional phosphocholine-binding miniantibodies could be increased in E. coli fermentations by the combination of the following approaches: Firstly, miniantibody mutants with amino acid exchanges in the VH chain leading to improved folding were expressed. Secondly, the expression vector was stabilized by an efficient suicide system to prevent plasmid loss. Thirdly, the cells were grown to high cell densities in a stirred tank reactor.
Kujau
M J
MJ
Hans-Knöll-Institut für Naturstoff-Forschung Jena, Dept. Applied Microbiology, Germany. mkujau@pmail.hki-jena.de
Riesenberg
D
D
D019943:Amino Acid Substitution / Q000235:genetics; D000907:Antibodies, Bacterial / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D016368:Frameshift Mutation; D005822:Genetic Vectors / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D010767:Phosphorylcholine / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D013997:Time Factors
D000907:Antibodies, Bacterial; D007143:Immunoglobulin Heavy Chains; D011994:Recombinant Proteins; D010767:Phosphorylcholine