Co-operative effects of protein engineering and vector optimization on high yield expression of functional bivalent miniantibodies in Escherichia coli

M J Kujau; D Riesenberg

doi:10.1016/S0944-5013(99)80031-7

Co-operative effects of protein engineering and vector optimization on high yield expression of functional bivalent miniantibodies in Escherichia coli

Microbiol Res. 1999 May;154(1):27-34. doi: 10.1016/S0944-5013(99)80031-7.

Authors

M J Kujau¹, D Riesenberg

Affiliation

¹ Hans-Knöll-Institut für Naturstoff-Forschung Jena, Dept. Applied Microbiology, Germany. [email protected]

PMID: 10356794
DOI: 10.1016/S0944-5013(99)80031-7

Abstract

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.

MeSH terms

Amino Acid Substitution / genetics
Antibodies, Bacterial* / genetics
Antibodies, Bacterial* / metabolism
Escherichia coli / genetics
Escherichia coli / immunology*
Escherichia coli / metabolism
Frameshift Mutation*
Genetic Vectors / genetics*
Immunoglobulin Heavy Chains / genetics
Phosphorylcholine / metabolism
Protein Engineering / methods*
Recombinant Proteins / biosynthesis
Time Factors

Substances

Antibodies, Bacterial
Immunoglobulin Heavy Chains
Recombinant Proteins
Phosphorylcholine