RT info:eu-repo/semantics/article
T1 A single-chain antibody fragment is functionally expressed in the cytoplasm of both Escherichia coli and transgenic plants
A1 Tavladoraki, Paraskevi
A1 Girotti ., Alessandra
A1 Donini, Marcello
A1 Arias Vallejo, Francisco Javier
A1 Mancini, Camillo
A1 Veronica Morea
A1 Roberta Chiaraluce
A1 Valerio Consalvi
A1 Eugenio Benvenuto
K1 Intrabody, scFv,
K1 (ScFv anticuerpos intracelulares, estabilidad proteinas
AB Despite the well-known crucial role of intradomain disulfide bridges for immunoglobulin folding and stability, thesingle-chain variable fragment of the anti-viral antibody F8 is functionally expressed when targeted to the reducingenvironment of the plant cytoplasm. We show here that this antibody fragment is also functionally expressed in thecytoplasm of Escherichia coli. A gel shift assay revealed that the single-chain variable fragment (scFv) accumulatingin the plant and bacterial cytoplasm bears free sulfhydryl groups. Guanidinium chloride denaturation/renaturationstudies indicated that refolding occurs even in a reducing environment, producing a functional molecule with thesame spectral properties of the native scFv(F8). Taken together, these results suggest that folding and functionality ofthis antibody fragment are not prevented in a reducing environment. This antibody fragment could thereforerepresent a suitable framework for engineering recombinant antibodies to be targeted to the cytoplasm.
PB Wiley
SN 0014-2956
YR 1999
FD 1999
LK https://uvadoc.uva.es/handle/10324/74194
UL https://uvadoc.uva.es/handle/10324/74194
LA eng
NO European Journal of Biochemistry (FEBS Journal). 262, 617-624
NO Producción Científica
DS UVaDOC
RD 22-ene-2025