Complete immunoglobulin molecules cannot be produced in Escherichia coli, but different strategies have been developed to express antibody fragments like single-chain antibodies (scFvs), disulfide stabilized antibodies (dsFvs), Fab fragments, bispecific antibodies (bsAbs) or various immunotoxins (overview in Breitling and Dübel, 1999). First, antibody fragments can be deposed as cytoplasmic inclusion bodies followed by refolding in vitro. This application is usually employed if the functional molecule is expected to kill the producing cell, e. g. for immunotoxins. Due to the general low folding efficiency, an alternative strategy has been developed which better imitates the folding conditions of antibodies in eucaryotic cells. The secretion of the antibody fragments into the periplasmic space of E. coli permits their production as soluble and functional proteins with correctly formed intramolecular disulfide bonds. The yields are usually lower than those of inclusion body refolding, however, it is the method of choice for a rapid assessment of antigen binding activity and specificity.
- Antibody Fragment
- Bispecific Antibody
- Intramolecular Disulfide Bond
- Periplasmic Extract
- Cytoplasmic Inclusion Body
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Schmiedl, A., Dübel, S. (2001). Functional Minipreps of scFv-Antibody Fragments. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Lab Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-04605-0_18
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