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A single-chain antibody fragment is functionally expressed in the cytoplasm of both Escherichia coli and transgenic plants

TitoloA single-chain antibody fragment is functionally expressed in the cytoplasm of both Escherichia coli and transgenic plants
Tipo di pubblicazioneArticolo su Rivista peer-reviewed
Anno di Pubblicazione1999
AutoriTavladoraki, P., Girotti A., Donini Marcello, Arias F.J., Mancini C., Morea V., Chiaraluce R., Consalvi V., and Benvenuto Eugenio
RivistaEuropean Journal of Biochemistry
Volume262
Paginazione617 - 624
Data di pubblicazione1999
ISBN Number00142956 (ISSN)
Parole chiaveAntibody, article, Cysteine, cytoplasm, cytosol, disulfide bond, Disulfides, Escherichia coli, Genetically Modified, immunoglobulin variable region, Intrabody, nonhuman, oxidation reduction state, Plants, priority journal, protein analysis, Protein Denaturation, protein expression, Protein Folding, Protein Sorting Signals, protein stability, scFv fragment, transgenic plant
Abstract

Despite the well-known crucial role of intradomain disulfide bridges for immunoglobulin folding and stability, the single-chain variable fragment of the anti-viral antibody F8 is functionally expressed when targeted to the reducing environment of the plant cytoplasm. We show here that this antibody fragment is also functionally expressed in the cytoplasm of Escherichia coli. A gel shift assay revealed that the single-chain variable fragment (scFv) accumulating in the plant and bacterial cytoplasm bears free sulfhydryl groups. Guanidinium chloride denaturation/renaturation studies indicated that refolding occurs even in a reducing environment, producing a functional molecule with the same spectral properties of the native scFv(F8). Taken together, these results suggest that folding and functionality of this antibody fragment are not prevented in a reducing environment. This antibody fragment could therefore represent a suitable framework for engineering recombinant antibodies to be targeted to the cytoplasm.

Note

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Citation Key5365